Nitrogen ring linked deoxyuridine triphosphatase inhibitors

ABSTRACT

Provided herein are dUTPase inhibitors, compositions comprising such compounds and methods of using such compounds and compositions.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/462,173, filed on May 17, 2019, which is a national stage applicationunder 35 U.S.C. § 371 of International Application No.PCT/US2017/062909, filed on Nov. 21, 2017, which claims the benefit ofpriority of U.S. Provisional Patent Application No. 62/426,149, filed onNov. 23, 2016, and U.S. Provisional Patent Application No. 62/547,721,filed on Aug. 18, 2017, of which the contents of each are incorporatedby reference herein in their entirety.

BACKGROUND

Thymidylate metabolism is required for producing essential buildingblocks necessary to replicate DNA in dividing cells and has long been animportant therapeutic target for cornerstone cancer drugs. Drugstargeting this pathway such as 5-fluorouracil (5-FU) inhibit the enzymethymidylate synthase (TS) and are currently critical standard-of caretherapies. TS-targeted agents are used for the treatment of a variety ofcancers including colon, gastric, head and neck, breast, lung and bloodrelated malignancies among others. Grem, J. L., 5-Fluorouracil plusleucovorin in cancer therapy, in Principals and Practice of OncologyUpdate Series, J. De Vita, V. T., S. Hellman, and A. Rosenberg, Editors.1988, J. B. Lippincott: Philadelphia, Pa.

There are two classes of drugs that target the TS enzyme: thefluoropyrimidines and the antifolates. The fluoropyrimidines, 5-FU, S-1and capecitabine (Xeloda®), have wide use in the treatment ofgastrointestinal and breast cancers, while the antifolate pemetrexed(Alimta®) is currently used for the treatment of non-small cell lungcancer (NSCLC). Since the discovery of 5-FU over fifty years ago byCharles Heidelberger, the fluoropyrimidines remain one of the mostcommon and effective anticancer cancer drugs used worldwide. Due to thisfact, there is an abundance of clinical experience and insight into themechanism of action of these agents.

The TS inhibitor 5-fluorouracil (5 FU) remains the foundation of manyfirst and second line regimens in the treatment of colon cancer. Singleagent therapies including oxaliplatin, irinotecan, Erbitux and Avastin,demonstrate lowered activity in colon cancer compared to 5-FU. Inaddition to colon cancer, TS-inhibitory agents have demonstratedefficacy in several other solid tumor types. Standard of care nowincorporates 5-FU as the backbone drug in combination with oxaliplatinor irinotecan or another agent.

Deoxyuridine triphosphatase (“dUTPase”) is a ubiquitous enzyme that isessential for viability in both prokaryotic and eukaryotic organisms; asthe main regulator of dUTP pools, the expression of dUTPase could haveprofound effects on the utility of chemotherapeutics that inhibitthymidylate biosynthesis. Normally, dUTPase mediates a protective roleby limiting the expansion of dUTP pools and countering the cytotoxiceffect of uracil misincorporation. According to this model, elevatedlevels of dUTPase could prevent TS inhibitor-induced dUTP accumulationand induce drug resistance. It has been shown that dUTPase overexpression results in a significant decrease in dUTP accumulation andincreased resistance to drug treatment when compared to controls.

Chemotherapeutic agents that target de novo thymidylate metabolism arecritical for the treatment of a variety of solid tumors, howeverclinical efficacy is often hindered by drug resistance. Becauseresistance to these agents is a common occurrence, the identificationand exploitation of novel determinants of drug sensitivity within thispathway of proven therapeutic utility is important. As disclosed byLadner et al. in U.S. Patent Publ. No. US 2011/0212467, the dUTPaseenzyme and the uracil-DNA misincorporation pathway can play a drivingrole in mediating cytotoxicity to TS-directed chemotherapies.

For example, nearly half of cancer patients do not benefit from5-FU-based treatment due to intrinsic or acquired drug resistance. Dueto this fact, there is a critical need to overcome the fundamentalchallenge of drug resistance and provide new therapeutic strategies toimprove patient outcome. This disclosure satisfies this need andprovides related advantages as well.

SUMMARY

In some aspects, this disclosure provides compounds, compositions andmethods that inhibit dUTPase when used alone or in combination with atleast one dUTPase-directed chemotherapy. In some aspects, thisdisclosure provides compounds, compositions and methods for treatingcancer, killing cancer cells, and/or inhibiting cancer cell growth whenused in combination with at least one TS-directed chemotherapy.Compounds of this class include, without limitation, the followingcompounds of formulas (I).

In one aspect, provided herein is a compound of Formula (IA):

or a tautomer thereof; or a prodrug of each thereof; or a deuteriumisotope of each of the above wherein up to 10, preferably up to 6, morepreferably up to 3 hydrogen atoms that are attached to one or morecarbon atoms are replaced with deuterium(s); or a pharmaceuticallyacceptable salt of each of the foregoing; or a pharmaceuticallyacceptable solvate of each of the above mentioned,

wherein

A is a uracil isostere, which is an optionally substituted, preferablyplanar or substantially planar (a substantially planar ring contains, 3or more SP² hybridized carbon and/or nitrogen atoms), ring comprising1-3 ring nitrogen atoms, wherein the ring is substituted with 1-3 groupsselected from keto or oxo, hydroxy, and alkoxy, and wherein the ring isa 6 membered monocyclic or a 9-10 membered bicyclic ring; or

A is an optionally substituted 5-membered heterocyclyl containing a—C(O)NZC(O)— moiety, a —C(O)OC(O) moiety, a —C(O)CR¹⁰C(O) moiety, or a—C(O)NR¹⁰C(O) moiety; or

A is a 5-membered heteroaryl or a 5-membered substantially planarheterocyclyl (i.e., a heterocyclyl wherein at least 3 or at least 4atoms can stably be in a same plane) substituted at 1,3 positions withsubstituents selected from halo, optionally substituted hydroxy, andoptionally substituted —SH groups, preferably two fluoros, wherein the5-membered heteroaryl or substantially planar heterocyclyl is furtheroptionally substituted; or

A is:

wherein X²⁰⁰ is N or CH; or

A is:

wherein A′ is an optionally substituted 4-7 membered cycloalkyl or anoptionally substituted 4-7 membered heterocyclyl; or

A is:

wherein A″ is an optionally substituted 5-7 membered cycloalkyl, anoptionally substituted 5-7 membered heterocyclyl, an optionallysubstituted 6-10 membered aryl, or an optionally substituted 6-10membered heteroaryl; or

A is

preferably:

or

A is

each R¹⁰ independently is hydrogen, an optionally substituted C₁-C₁₀alkoxy, or an optionally substituted C₁-C₁₀ alkyl, preferably R¹⁰ ishydrogen;

each R³⁰ independently is hydrogen; an optionally substituted C₁-C₁₀alkoxy; optionally substituted amino, such as —NH₂ or a mono ordi-substituted form thereof; an optionally substituted C₁-C₁₀ alkyl;optionally substituted hydroxy; a prodrug moiety, or Z; or A and L¹,preferably, R³⁰, wherein R³⁰ is attached to an atom that is adjacent tothe atom attached to L¹, and L¹ together with the atoms they areattached to form a 5-7 membered ring;

L¹ is a linker having 2-8 chain atoms selected from C, N, O, S, and/orP, wherein the linker is optionally substituted;

L² is —C(O)— or —S(O)₂—;

the nitrogen containing ring attached to L² and L³ is optionallysubstituted;

is an optionally substituted 4-6 membered heterocyclyl or an optionallysubstituted 5-6 membered heteroaryl group containing at least one ringnitrogen atom;

L³ is a bond or —C(R³⁰⁰)₂—;

each R³⁰⁰ independently is a hydrogen, hydroxyl, an optionallysubstituted C₁-C₆ alkyl, or an optionally substituted C₆-C₁₀ aryl, suchas without limitation, phenyl;

each R⁵⁰ independently is hydrogen, an optionally substituted C₁-C₆alkyl, an optionally substituted C₂-C₆ heteroalkyl, an optionallysubstituted C₂-C₆ alkenyl, an optionally substituted C₃-C₆heteroalkenyl, an optionally substituted C₂-C₆ alkynyl, an optionallysubstituted C₃-C₆ heteroalkynyl, or Z;

Z is

each R⁵¹ and R⁵² independently is hydrogen or an optionally substitutedC₁-C₁₀ alkyl;

X is an optionally substituted hydroxy group, an optionally substitutedNH₂ group, or an optionally substituted SH group; and

B is an optionally substituted 6-10 membered aryl; an optionallysubstituted 5-15 membered heteroaryl; an optionally substituted 4-15membered heterocyclyl; or an optionally substituted 3-15 memberedcycloalkyl, if cycloalkyl, then preferably at least a 4 membered, ormore preferably a 5-10 membered cycloalkyl; or a B substituent togetherwith L² or L³, form a 5-7 membered cycloalkyl or a heterocyclyl.

In one embodiment, the compound provided herein is of formula (I):

wherein the variables are defined as in formula (IA).

In one embodiment,

is

In one embodiment,

is

In one embodiment,

is

In some embodiments, the compound provided herein is a prodrug. As usedherein, “prodrug” refers to a compound that, after administration, ismetabolized or otherwise converted to a biologically active or moreactive compound (or drug) with respect to at least one property. Aprodrug, relative to the drug, is modified chemically in a manner thatrenders it, relative to the drug, less active or inactive, but thechemical modification is such that the corresponding drug is generatedby metabolic or other biological processes after the prodrug isadministered. A prodrug may have, relative to the active drug, alteredmetabolic stability or transport characteristics, fewer side effects orlower toxicity, or improved flavor (for example, see the referenceNogrady, 1985, Medicinal Chemistry A Biochemical Approach, OxfordUniversity Press, New York, pages 388-392, incorporated herein byreference). A prodrug may be synthesized using reactants other than thecorresponding drug. Examples of prodrugs and methods of making them arealso provided in US Patent Application Publication No. 20160024127,which is incorporated herein in its entirety by reference.

In some embodiments, the compound provided herein contains one or moredeuterium. Examples of a deuterium containing compound provided herein,wherein up to 10, preferably up to 6, more preferably up to 3 hydrogenatoms that are attached to carbon atoms are replaced with a deuterium,include, without limitation: a compound where a methyl group isconverted to —CH₂D, —CHD₂, or —CD₃; a compound where a methylene groupis converted to a —CHD- or —CD₂-, a phenyl ring where one or morehydrogen atoms are replaced with deuterium atoms, etc.

In some embodiments, A is an optionally substituted 5-memberedheterocyclyl containing a —C(O)NZC(O)— moiety. In some embodiments, A isan optionally substituted 5-membered heterocyclyl containing a—C(O)OC(O) moiety. In some embodiments, A is an optionally substituted5-membered heterocyclyl containing a —C(O)CR¹⁰C(O) moiety. In someembodiments, A is an optionally substituted 5-membered heterocyclylcontaining a —C(O)NR¹⁰C(O) moiety.

In some embodiments, R¹⁰ is hydrogen. In some embodiments, R¹⁰ is anoptionally substituted C₁-C₁₀ alkoxy. In some embodiments, R¹⁰ is anoptionally substituted C₁-C₁₀ alkyl.

In some embodiments, A is a 5-membered heteroaryl substituted at 1,3positions with substituents selected from halo, optionally substitutedhydroxy, and optionally substituted —SH groups, preferably two fluoros,wherein the 5-membered heteroaryl is further optionally substituted. Insome embodiments, A is a 5-membered heteroaryl substituted at 1,3positions with halo, wherein the 5-membered heteroaryl is furtheroptionally substituted. In some embodiments, the 5-membered heteroarylis substituted at 1,3 positions with two fluoros, wherein the 5-memberedheteroaryl is further optionally substituted. In some embodiments, A isa 5-membered heteroaryl substituted at 1,3 positions with optionallysubstituted hydroxy, wherein the 5-membered heteroaryl is furtheroptionally substituted. In some embodiments, A is a 5-memberedheteroaryl substituted at 1,3 positions with optionally substituted —SHgroups, wherein the 5-membered heteroaryl is further optionallysubstituted.

Non-limiting and illustrative examples of a 5-membered heteroarylsubstituted at 1,3 positions with substituents selected from halo,optionally substituted hydroxy, optionally substituted —SH groupsinclude, without limitation:

such as

where Y¹⁰ and Y¹¹ independently are selected from a halo, preferablychloro or fluoro, hydroxy, —SH, substituted hydroxy, and substituted—SH; Z²⁰-Z²² are independently selected from optionally substituted CH,optionally substituted NH, N, S, SO₂, SO, and O, provided that thecombination of Z²⁰-Z²² provides a planar valence matched heteroaryl or atautomer thereof; and each Z²³ independently is CH or N.

In some embodiments, Y¹⁰ is a halo. In some embodiments, Y¹⁰ is achloro. In some embodiments, Y¹⁰ is a fluoro. In some embodiments, Y¹⁰is hydroxy. In some embodiments, Y¹⁰ is —SH. In some embodiments, Y¹⁰ isa substituted hydroxy. In some embodiments, Y¹⁰ is a substituted —SH.

In some embodiments, Y¹¹ is a halo. In some embodiments, Y¹¹ is achloro. In some embodiments, Y¹¹ is a fluoro. In some embodiments, Y¹¹is hydroxy. In some embodiments, Y¹¹ is —SH. In some embodiments, Y¹¹ isa substituted hydroxy. In some embodiments, Y¹¹ is a substituted —SH.

In some embodiments, Z²⁰ is an optionally substituted CH. In someembodiments, Z²⁰ is an optionally substituted NH. In some embodiments,Z²⁰ is N. In some embodiments, Z²⁰ is S. In some embodiments, Z²⁰ isSO₂. In some embodiments, Z²⁰ is SO. In some embodiments, Z²⁰ is O.

In some embodiments, Z²¹ is an optionally substituted CH. In someembodiments, Z²¹ is an optionally substituted NH. In some embodiments,Z²¹ is N. In some embodiments, Z²¹ is S. In some embodiments, Z²¹ isSO₂. In some embodiments, Z²¹ is SO. In some embodiments, Z²¹ is O.

In some embodiments, Z²² is an optionally substituted CH. In someembodiments, Z²² is an optionally substituted NH. In some embodiments,Z²² is N. In some embodiments, Z²² is S. In some embodiments, Z²² isSO₂. In some embodiments, Z²² is SO. In some embodiments, Z²² is O.

In some embodiments, Z²³ is an optionally substituted CH. In someembodiments, Z²³ is N.

In some embodiments, A is a 5-membered substantially planar heterocyclyl(i.e., a heterocyclyl wherein at least 3 or at least 4 atoms can stablybe in a same plane) substituted at 1,3 positions with substituentsselected from halo, optionally substituted hydroxy, and optionallysubstituted —SH groups, preferably two fluoros, wherein the 5-memberedsubstantially planar heterocyclyl is further optionally substituted. Insome embodiments, A is a 5-membered substantially planar heterocyclylsubstituted at 1,3 positions with halo, wherein the 5-memberedsubstantially planar heterocyclyl is further optionally substituted. Insome embodiments, the 5-membered substantially planar heterocyclyl issubstituted at 1,3 positions with two fluoros, wherein the 5-memberedsubstantially planar heterocyclyl is further optionally substituted. Insome embodiments, A is a 5-membered substantially planar heterocyclylsubstituted at 1,3 positions with optionally substituted hydroxy,wherein the 5-membered substantially planar heterocyclyl is furtheroptionally substituted. In some embodiments, A is a 5-memberedsubstantially planar heterocyclyl substituted at 1,3 positions withoptionally substituted —SH groups, wherein the 5-membered substantiallyplanar heterocyclyl is further optionally substituted.

Examples of a 5-membered substantially planar heterocyclyl substitutedat 1,3 positions with halo, optionally substituted hydroxy, andoptionally substituted —SH groups, have similar structures as thecorresponding 5-membered heteroaryl except that the 5-membered ring isnot an aromatic ring.

In some embodiments, A is selected from the group consisting of:

wherein each R³⁰ is as defined above.

In some embodiments, A is selected from the group consisting of:

wherein each R³⁰ is as defined above.

In some embodiments, A is a uracil isostere, which is an optionallysubstituted, preferably planar or substantially planar (a substantiallyplanar ring contains, 3 or more SP² hybridized carbon and/or nitrogenatoms), ring comprising 1-3 ring nitrogen atoms, wherein the ring issubstituted with 1-3 groups selected from keto or oxo, hydroxy, andalkoxy, and wherein the ring is a 6 membered monocyclic or a 9-10membered bicyclic ring.

In some embodiments, A is:

In some embodiments, A is:

In some embodiments, A is:

In some embodiments, A is:

In some embodiments, A is:

In some embodiments, A is:

In some embodiments, A is:

In some embodiments, A is:

In some embodiments, A is:

In some embodiments, A is:

In some embodiments, A is:

In some embodiments, A is:

In some embodiments, A is:

In some embodiments, A is:

In some embodiments, A is:

In some embodiments, A is:

In some embodiments, R³⁰ is hydrogen. In some embodiments, R³⁰ is anoptionally substituted C₁-C₁₀ alkoxy. In some embodiments, R³⁰ isoptionally substituted amino, such as —NH₂ or a mono or di-substitutedform thereof. In some embodiments, R³⁰ is an optionally substitutedC₁-C₁₀ alkyl. In some embodiments, R³⁰ is an optionally substitutedhydroxy. In some embodiments, R³⁰ is a prodrug moiety. Non-limiting andillustrative prodrug moieties include formyl ethers, and formyl estersas disclosed herein. In some embodiments, R³⁰ is Z.

Illustrative and non-limiting examples of R³⁰ include a substitutedhydroxy or —CH₂OC(O)R⁸⁰, wherein R⁸⁰ is H or an optionally substitutedC₁-C₁₀ alkyl. In some embodiments, R⁸⁰ is hydrogen. In some embodiments,R⁸⁰ is an optionally substituted C₁-C₁₀ alkyl.

In some embodiments, A and L¹, preferably, R³⁰ and L¹ together with theatoms they are attached to form a 5-7 membered ring.

In some embodiments, A is selected from the group consisting of:

In some embodiments, A is

In some embodiments, A is

In some embodiments, A is

In some embodiments, A is

In some embodiments, A is

In some embodiments, A is

The A moieties disclosed herein including herein above, can, in someembodiments, be further substituted with 1-3, preferably 1-2, morepreferably, 1 R¹⁰ substituent as provided herein. In some embodiments,where R¹⁰ and L¹ are joined to adjacent atoms (i.e., atoms having a 1,2positional relation), R¹⁰ and a portion of L¹, together with theintervening atoms can form a 5-6 membered, optionally substitutedcycloalkyl or heterocyclyl ring.

In some embodiments, L¹ is a linker having 2-8 chain atoms selected fromC, N, O, S, and/or P, wherein the linker is optionally substituted. Invarious embodiments, L¹ having 2-8 chain atoms selected from C, N, O, S,and/or P can be: alkylene, alkenylene, alkynylene, wherein one or morecarbon atoms are replaced with O, S, SO, SO₂, optionally substituted NH,

moieties where R^(Q) is H or C₁-C₆ alkyl optionally substituted —CO—NH—,optionally substituted —SO₂—NH—, optionally substituted —P(O)(OH)—,optionally substituted phosphoramide and optionally substitutedphosporamidate, (such as —P(O)NH₂—, —P(O)(OH)NH—, etc.), optionallysubstituted oligoethylene glycol, optionally substituted oligoethanolamine, and the likes, as will be apparent to the skilled artisanbased on the disclosure provided herein.

In some embodiments, L¹ is —(CH₂)_(q)—. In some embodiments, one or morehydrogens are optionally substituted with C₁-C₃ alkyl. In someembodiments, at least two or more geminal hydrogens are optionallysubstituted with an optionally substituted 3-5 membered heterocyclyl. Insome embodiments, at least two or more geminal hydrogens are optionallysubstituted with an optionally substituted 3-5 membered cycloalkyl. Insome embodiments, the optionally substituted 3-5 membered cycloalkyl isan optionally substituted cyclopropano. In some embodiments, theoptionally substituted 3-5 membered cycloalkyl is an optionallysubstituted cyclobutano. In some embodiments, the optionally substituted3-5 membered cycloalkyl is an optionally substituted cyclopentano. Insome embodiments, the optionally substituted 3-5 membered cycloalkyl isan optionally substituted tetrahydrofurano.

In some embodiments, q is 3. In some embodiments, q is 4. In someembodiments, q is 5. In some embodiments, q is 6. In some embodiments, qis 7. In some embodiments, q is 8.

In some embodiments, L¹ is:

In some related embodiments, one or more hydrogens are optionallysubstituted with C₁-C₃ alkyl. In some embodiments, at least two or moregeminal hydrogens are optionally substituted with an optionallysubstituted 3-5 membered heterocyclyl. In some embodiments, at least twoor more geminal hydrogens are optionally substituted with an optionallysubstituted 3-5 membered cycloalkyl. In some embodiments, the optionallysubstituted 3-5 membered cycloalkyl is an optionally substitutedcyclopropano. In some embodiments, the optionally substituted 3-5membered cycloalkyl is an optionally substituted cyclobutano. In someembodiments, the optionally substituted 3-5 membered cycloalkyl is anoptionally substituted cyclopentano. In some embodiments, the optionallysubstituted 3-5 membered cycloalkyl is an optionally substitutedtetrahydrofurano.

In some embodiments, p is 0. In some embodiments, p is 1. In someembodiments, p is 2. In some embodiments, p is 3. In some embodiments, pis 4. In some embodiments, p is 5.

In some embodiments, z is 0. In some embodiments, z is 1. In someembodiments, z is 2. In some embodiments, z is 3. In some embodiments, zis 4. In some embodiments, z is 5.

In some embodiments, L¹ is —(CH₂)_(m)—X¹⁵—(CH₂)_(n)—. In someembodiments, one or more hydrogens are optionally substituted with C₁-C₃alkyl. In some embodiments, at least two or more geminal hydrogens areoptionally substituted with an optionally substituted 3-5 memberedheterocyclyl. In some embodiments, at least two or more geminalhydrogens are optionally substituted with an optionally substituted 3-5membered cycloalkyl. In some embodiments, the optionally substituted 3-5membered cycloalkyl is an optionally substituted cyclopropano. In someembodiments, the optionally substituted 3-5 membered cycloalkyl is anoptionally substituted cyclobutano. In some embodiments, the optionallysubstituted 3-5 membered cycloalkyl is an optionally substitutedcyclopentano. In some embodiments, the optionally substituted 3-5membered cycloalkyl is an optionally substituted tetrahydrofurano.

In some embodiments, m is 0. In some embodiments, m is 1. In someembodiments, m is 2. In some embodiments, m is 3.

In some embodiments, n is 0. In some embodiments, n is 1. In someembodiments, n is 2. In some embodiments, n is 3. In some embodiments, nis 4. In some embodiments, n is 5. In some embodiments, n is 6. In someembodiments, n is 7.

In some embodiments, X¹⁵ is NR⁴⁰. In some embodiments, X¹⁵ is—NR⁴⁰(+)-O(−). In some embodiments, R⁴⁰ is H. In some embodiments, R⁴⁰is C₁-C₃ alkyl. In some embodiments, X¹⁵ is O. In some embodiments, X¹⁵is S. In some embodiments, X¹⁵ is SO. In some embodiments, X¹⁵ is SO₂.

In some embodiments, L¹ is:

where X¹⁵ is defined as above. In some related embodiments, one or morehydrogens are optionally substituted with C₁-C₃ alkyl. In someembodiments, at least two or more geminal hydrogens are optionallysubstituted with an optionally substituted 3-5 membered heterocyclyl. Insome embodiments, at least two or more geminal hydrogens are optionallysubstituted with an optionally substituted 3-5 membered cycloalkyl. Insome embodiments, the optionally substituted 3-5 membered cycloalkyl isan optionally substituted cyclopropano. In some embodiments, theoptionally substituted 3-5 membered cycloalkyl is an optionallysubstituted cyclobutano. In some embodiments, the optionally substituted3-5 membered cycloalkyl is an optionally substituted cyclopentano. Insome embodiments, the optionally substituted 3-5 membered cycloalkyl isan optionally substituted tetrahydrofurano.

In some embodiments, o is 0. In some embodiments, o is 1. In someembodiments, o is 2. In some embodiments, o is 3.

In some embodiments, r is 1. In some embodiments, r is 2. In someembodiments, r is 3.

In some embodiments, s is 0. In some embodiments, s is 1. In someembodiments, s is 2. In some embodiments, s is 3. In some embodiments, sis 4.

In some embodiments, L′ is selected from the group consisting of:

In some related embodiments, 1-5, preferably, 1-3 hydrogen atoms of theL¹ are optionally substituted, preferred substituents including withoutlimitation, C₁-C₆ alkyl optionally substituted with 1-3 halo, such asfluoro, and/or C₁-C₆ alkoxy; optionally substituted C₁-C₆ alkoxy; andhalo, preferably fluoro, wherein the left side of the moieties areattached to A and wherein R⁷⁰ is a hydrogen or an optionally substitutedC₁-C₁₀ alkyl. In some embodiments, L¹ is optionally substituted wherein1-5 hydrogen atoms are optionally substituted. In some embodiments, L¹is optionally substituted wherein 1-3 hydrogen atoms are optionallysubstituted. In some embodiments, substituents include withoutlimitation C₁-C₆ alkyl optionally substituted with 1-3 halo, such asfluoro. In some embodiments, substituents include without limitationC₁-C₆ alkyl optionally substituted with C₁-C₆ alkoxy. In someembodiments, substituents include without limitation an optionallysubstituted C₁-C₆ alkoxy. In some embodiments, substituents includewithout limitation a halo. In some embodiments, substituents include afluoro.

In some embodiments, L¹ is:

or an optionally substituted version of each thereof wherein 1-5,preferably, 1-3 hydrogen atoms are optionally substituted, preferredsubstituents including without limitation, C₁-C₆ alkyl optionallysubstituted with 1-3 halo, such as fluoro, and/or C₁-C₆ alkoxy;optionally substituted C₁-C₆ alkoxy; and halo, preferably fluoro,wherein the left side of the moieties are attached to A.

In some embodiments, L¹ is:

In some embodiments, L¹ is:

In some embodiments, L¹ is:

In some embodiments, L¹ is optionally substituted wherein 1-5 hydrogenatoms are optionally substituted. In some embodiments, L¹ is optionallysubstituted wherein 1-3 hydrogen atoms are optionally substituted. Insome embodiments, substituents include without limitation C₁-C₆ alkyloptionally substituted with 1-3 halo, such as fluoro. In someembodiments, substituents include without limitation C₁-C₆ alkyloptionally substituted with C₁-C₆ alkoxy. In some embodiments,substituents include without limitation an optionally substituted C₁-C₆alkoxy. In some embodiments, substituents include without limitation ahalo. In some embodiments, substituents include a fluoro.

In some embodiments, L² is —C(O)—. In some embodiments, L² is —S(O)₂—.

In some embodiments, Z is

wherein each R⁵¹ and R⁵² independently is hydrogen or an optionallysubstituted C₁-C₁₀ alkyl and X is an optionally substituted hydroxygroup, an optionally substituted NH₂ group, or an optionally substitutedSH group.

In some embodiments, R⁵¹ is hydrogen. In some embodiments, R⁵¹ is anoptionally substituted C₁-C₁₀ alkyl. In some embodiments, R⁵² ishydrogen. In some embodiments, R⁵² is an optionally substituted C₁-C₁₀alkyl.

In some embodiments, X is an optionally substituted hydroxy group. Insome embodiments, X is an optionally substituted NH₂ group. In someembodiments, X is an optionally substituted SH group.

As used herein, an optionally substituted hydroxy group refers towithout limitation alkylated, arylated, cycloalkylated,heterocyclylated, acylated, carboxylated (i.e., generating a carbonate,carbamate, a thiocarbonate, a thiacarbamate containing alkyl, aryl,heteroaryl, and/or heterocyclyl, and such other moieties),phosphorylated, phosphonylated, sulfonylated, forms of a hydroxy group,as would be apparent to the skilled artisan in view of this disclosure.

As used herein, an optionally substituted NH₂ group refers to withoutlimitation alkylated, arylated, cycloalkylated, heterocyclylated,acylated, carboxylated (i.e., generating a carbonate, carbamate, athiocarbonate, a thiacarbamate containing alkyl, aryl, heteroaryl,and/or heterocyclyl, and such other moieties), phosphorylated,phosphonylated, sulfonylated, forms of a NH₂ group, as would be apparentto the skilled artisan in view of this disclosure.

As used herein, an optionally substituted SH group refers to withoutlimitation alkylated, arylated, cycloalkylated, heterocyclylated,acylated, carboxylated (i.e., generating a carbonate, carbamate, athiocarbonate, a thiacarbamate containing alkyl, aryl, heteroaryl,and/or heterocyclyl, and such other moieties), phosphorylated,phosphonylated, sulfonylated, forms of a —SH group, as would be apparentto the skilled artisan in view of this disclosure.

In some embodiments, L³ is a bond. In some embodiments, L³ is—C(R³⁰⁰)₂—.

In some embodiments, each R³⁰⁰ independently is a hydrogen, hydroxyl, oran optionally substituted phenyl. In some embodiments, each R³⁰⁰independently is a hydrogen. In some embodiments, each R³⁰⁰independently is a hydroxyl. In some embodiments, each R³⁰⁰independently is an optionally substituted phenyl.

In some embodiments, one R³⁰⁰ is hydroxyl and the second R³⁰⁰ is anoptionally substituted phenyl. In some embodiments, one R³⁰⁰ is hydrogenand the second R³⁰⁰ is an optionally substituted phenyl. In someembodiments, one R³⁰⁰ is a phenyl, wherein the phenyl is notsubstituted.

As used herein, a hydantoin moiety refers to:

wherein R³⁰ is as defined above.

In some embodiments, a hydantoin moiety is:

In some embodiments, B is an optionally substituted 6-10 membered aryl.In some embodiments, B is an optionally substituted 5-15 memberedheteroaryl. In some embodiments, B is an optionally substituted 4-15membered heterocyclyl. In some embodiments, B is an optionallysubstituted 3-15 membered cycloalkyl. In some embodiments, if B is a3-15 membered cycloalkyl, then B is at least a 4 membered cycloalkyl. Insome embodiments, if B is a 3-15 membered cycloalkyl, then B is a 5-10membered cycloalkyl.

In some embodiments, B is selected from the group consisting of:

wherein

each R⁶ independently is hydrogen, an optionally substituted C₁-C₆alkoxy, or halo;

each R⁷ independently is an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₂-C₆ alkenyl, an optionally substituted C₂-C₆alkynyl, an optionally substituted C₃-C₈ cycloalkyl, an optionallysubstituted C₃-C₁₀ heteroaryl, an optionally substituted C₃-C₁₀heterocyclyl, or an optionally substituted C₆-C₁₀ aryl such asoptionally substituted phenyl; or

R⁶ and R⁷ together with the atoms they are attached to form anoptionally substituted 5-7 membered ring; or 2 R⁶ groups together withthe atoms they are attached to form an optionally substituted 5-7membered ring;

each R⁶¹ and R⁶² is independently N or CH, provided that at least one ofR⁶¹ and R⁶² is N,

each R⁶³ is independently NR⁹⁰, S, or O;

each R⁶⁴ is independently N or CH; and

each R⁹⁰ is independently hydrogen or R⁷,

and wherein one or more hydrogen atoms on the 5 and 6 membered aryl orheteroaryl rings shown above can be further optionally substituted.

In some embodiments, B is:

In some embodiments, B is:

In some embodiments, B is:

In some embodiments, B is:

In some embodiments, B is:

In some embodiments, B is:

In some embodiments, B is:

In some embodiments, B is:

In some embodiments, B is:

In some embodiments, B is:

In some embodiments, B is:

In some embodiments, B is:

In some embodiments, R⁶ is hydrogen. In some embodiments, R⁶ is anoptionally substituted C₁-C₆ alkoxy. In some embodiments, R⁶ is halo.

In some embodiments, R⁷ is an optionally substituted C₁-C₆ alkyl. Insome embodiments, R⁷ is an optionally substituted C₂-C₆ alkenyl. In someembodiments, R⁷ is an optionally substituted C₂-C₆ alkynyl. In someembodiments, R⁷ is an optionally substituted C₃-C₈ cycloalkyl. In someembodiments, R⁷ is an optionally substituted C₃-C₁₀ heteroaryl. In someembodiments, R⁷ is an optionally substituted C₃-C₁₀ heterocyclyl. Insome embodiments, R⁷ is an optionally substituted C₆-C₁₀ aryl. In someembodiments, the optionally substituted C₆-C₁₀ aryl is an optionallysubstituted phenyl.

In some embodiments, R⁶ and R⁷ together with the atoms they are attachedto form an optionally substituted 5-7 membered ring. In someembodiments, 2 R⁶ groups together with the atoms they are attached toform an optionally substituted 5-7 membered ring.

In some embodiments, one of R⁶¹ and R⁶² is N. In some embodiments, boththe R⁶¹ and R⁶² are N.

In some embodiments, R⁶³ is NR⁹⁰. In some embodiments, R⁶³ is S. In someembodiments, R⁶³ is O.

In some embodiments, R⁶⁴ is N. In some embodiments, R⁶⁴ is CH.

In some embodiments, R⁹⁰ is hydrogen. In some embodiments, R⁹⁰ is R⁷.

In some embodiments, B is

wherein

each R¹-R³ independently is H, halo, an optionally substituted C₁-C₆alkyl, an optionally substituted 4-15 membered heterocyclyl, or —OR²⁰or, if two of R¹-R³ are on adjacent carbon atoms, then two suchsubstituents together with the atoms they are attached to form anoptionally substituted 5-7 membered ring;

R²⁰ is (CH₂)_(w)—R²¹, an optionally substituted C₃-C₆ cycloalkyl, or anoptionally substituted C₁-C₆ alkyl;

R²¹ is an optionally substituted C₁-C₁₀ alkyl, an optionally substitutedC₂-C₁₀ alkenyl, an optionally substituted C₂-C₁₀ alkynyl, an optionallysubstituted C₃-C₆ cycloalkyl, optionally substituted phenyl, optionallysubstituted 5-15 membered heteroaryl, an optionally substituted 4-15membered heterocyclyl, or

wherein each R²²-R²⁴ independently is an optionally substituted C₁-C₃alkyl or hydroxy or two of R²²-R²⁴ together with the carbon atoms theyare attached to form a 3-7 membered, preferably a 3-5 membered, or a 5-7membered ring; and

w is 1, 2, 3, 4, or 5.

In some embodiments, R¹ is H. In some embodiments, R¹ is halo. In someembodiments, R¹ is an optionally substituted C₁-C₆ alkyl. In someembodiments, R¹ is H. In some embodiments, R¹ is an optionallysubstituted 4-15 membered heterocyclyl. In some embodiments, R¹ is—OR²⁰.

In some embodiments, R² is H. In some embodiments, R² is halo. In someembodiments, R² is an optionally substituted C₁-C₆ alkyl. In someembodiments, R² is H. In some embodiments, R² is an optionallysubstituted 4-15 membered heterocyclyl. In some embodiments, R² is—OR²⁰.

In some embodiments, R³ is H. In some embodiments, R³ is halo. In someembodiments, R³ is an optionally substituted C₁-C₆ alkyl. In someembodiments, R³ is H. In some embodiments, R³ is an optionallysubstituted 4-15 membered heterocyclyl. In some embodiments, R³ is—OR²⁰.

In some embodiments, if two of R¹-R³ are on adjacent carbon atoms, thentwo such substituents together with the atoms they are attached to forman optionally substituted 5-7 membered ring.

In some embodiments, R²⁰ is (CH₂)_(w)—R²¹. In some embodiments, R²⁰ isan optionally substituted C₃-C₆ cycloalkyl. In some embodiments, R²⁰ isan optionally substituted C₁-C₆ alkyl. In some embodiments, R²⁰ is anC₁-C₆ alkyl. In some embodiments, R²⁰ is an C₁-C₆ alkyl substituted with1-3 fluoro. In some embodiments, R²⁰ is an C₁-C₆ alkyl substituted with1-2, preferably, a single hydroxy.

In some embodiments, w is 1. In some embodiments, w is 2. In someembodiments, w is 3. In some embodiments, w is 4. In some embodiments, wis 5.

In some embodiments, R²¹ is a C₃-C₆ cycloalkyl. In some embodiments, R²¹is a C₃-C₆ cycloalkyl substituted with 1-3, preferably 1-2 substituents.In some embodiments, R²¹ is a cyclopropyl. In some embodiments, R²¹ is acyclopropyl substituted with 1-3, preferably 1-2 substituents. In someembodiments, R²¹ is a cyclobutyl. In some embodiments, R²¹ is acyclobutyl substituted with 1-3, preferably 1-2 substituents. In someembodiments, R²¹ is a cyclopentyl. In some embodiments, R²¹ is acyclopentyl substituted with 1-3, preferably 1-2 substituents. In someembodiments, R²¹ is an optionally substituted C₁-C₁₀ alkyl. In someembodiments, R²¹ is an optionally substituted C₂-C₁₀ alkenyl. In someembodiments, R²¹ is an optionally substituted C₂-C₁₀ alkynyl. In someembodiments, R²¹ is an optionally substituted 4-15 memberedheterocyclyl.

In some embodiments, R²¹ is

In some embodiments, R²² is an optionally substituted C₁-C₃ alkyl. Insome embodiments, R²² is hydroxy. In some embodiments, R²² is H.

In some embodiments, R²³ is an optionally substituted C₁-C₃ alkyl. Insome embodiments, R²³ is hydroxy.

In some embodiments, R²⁴ is an optionally substituted C₁-C₃ alkyl. Insome embodiments, R²⁴ is hydroxy.

In some embodiments, two of R²²-R²⁴ together with the carbon atoms theyare attached to form a 3-7 membered ring. In some embodiments, two ofR²²-R²⁴ together with the carbon atoms they are attached to form a 5-7membered ring. In some embodiments, the ring is optionally substitutedcycloalkyl. In some embodiments, the ring is optionally substitutedheterocyclyl.

In some embodiments, B is

wherein

R¹, R², and R³ are as defined above; or

R¹ and R² together with the atoms they are attached to form anoptionally substituted 5-7 membered ring; or

R² and R³ together with the atoms they are attached to form anoptionally substituted 5-7 membered ring.

In some embodiments, R¹ and R² together with the atoms they are attachedto form an optionally substituted 5-7 membered ring. In someembodiments, R² and R³ together with the atoms they are attached to forman optionally substituted 5-7 membered ring.

In some embodiments, wherein R¹ is H.

In some embodiments, R² is F. In some embodiments, R² is H.

In some embodiments, R³ is H. In some embodiments, R³ is —OR²⁰, whereinR²⁰ is as defined above.

In some embodiments, B is:

and wherein R²⁰ is as defined above.

In some embodiments, provided herein is a compound wherein A is

Y¹ is H or C₁-C₃ alkyl;

L¹ is an optionally substituted C₃-C₁₀ alkylene, further wherein atleast two geminal hydrogens are optionally substituted with cyclopropanoor cyclobutano; optionally substituted C₃-C₁₀ alkenylene, optionallysubstituted C₃-C₁₀ heteroalkylene, optionally substituted C₃-C₁₀heteroalkenylene, or -L¹¹-L¹²-L¹³-, wherein L¹¹ is attached to A and L¹¹is O, S, NR, C₁-C₂ alkylene, C₂ alkenylene, C₂ heteroalkylene, C₃heteroalkenylene, L¹² is arylene or heteroarylene, L¹³ is a bond or anoptionally substituted C₁-C₅ alkylene, and R is H or C₁-C₃ alkyl;

L² is —S(O)₂NH—, wherein the sulfur is attached to L¹ or —NHS(O)₂—,wherein the nitrogen is attached to L¹;

L³ is a bond or an optionally substituted C₁-C₆ alkylene;

B is

each R¹-R³ independently is H, F, Cl, C₁-C₃ alkyl, or —OR²⁰; or

R¹ and R² together with the atoms they are attached to form anoptionally substituted 5-7 membered ring; or

R² and R³ together with the atoms they are attached to form anoptionally substituted 5-7 membered ring;

R²⁰ is CH₂-R²¹; methyl optionally substituted with 2 or 3 fluorineatoms; C₃-C₆ cycloalkyl; or C₁-C₆ alkyl;

R²¹ is an optionally substituted C₃-C₆ cycloalkyl; an optionallysubstituted C₆-C₁₀ aryl; an optionally substituted 5-15 memberedheteroaryl; an optionally substituted 4-15 membered heterocyclyl; C₁-C₁₀alkyl, preferably branched C₃-C₁₀ alkyl optionally substituted with oneor more hydroxy or fluoro; C₃-C₆ cycloalkyl; or

wherein each R²²-R²⁴ independently is an optionally substituted C₁-C₃alkyl or hydroxy; or

two of R²²-R²⁴ together with the atoms they are attached to form anoptionally substituted 3-7 membered ring.

In some embodiments, Y¹ is H. In some embodiments, Y¹ is C₁-C₃ alkyl.

In some embodiments, L¹ is an optionally substituted C₃-C₁₀ alkylene,further wherein at least two geminal hydrogens are optionallysubstituted with cyclopropano or cyclobutano. In some embodiments, L¹ isan optionally substituted C₃-C₁₀ alkenylene. In some embodiments, L¹ isoptionally substituted C₃-C₁₀ heteroalkylene. In some embodiments, L¹ isoptionally substituted C₃-C₁₀ heteroalkenylene.

In some embodiments, L¹ is -L¹¹-L¹²-L¹³- wherein L¹¹ is attached to A.In some embodiments, L¹¹ is O. In some embodiments, L¹¹ is S. In someembodiments, L¹¹ is C₁-C₂ alkylene. In some embodiments, L¹¹ is C₂alkenylene. In some embodiments, L¹¹ is C₂ heteroalkylene. In someembodiments, L¹¹ is C₃ heteroalkenylene.

In some embodiments, L¹¹ is NR. In some embodiments, R is H. In someembodiments, R is C₁-C₃ alkyl.

In some embodiments, L¹² is arylene. In some embodiments, L¹² isheteroarylene.

In some embodiments, L¹³ is a bond. In some embodiments, L¹³ is anoptionally substituted C₁-C₅ alkylene.

In some embodiments, L² is —S(O)₂NH—, wherein the sulfur is attached toL¹ or —NHS(O)₂—, wherein the nitrogen is attached to L¹.

In some embodiments, L³ is a bond. In some embodiments, L³ is anoptionally substituted C₁-C₆ alkylene.

In some embodiments, R¹ is H. In some embodiments, R¹ is F. In someembodiments, R¹ is Cl. In some embodiments, R¹ is C₁-C₃ alkyl. In someembodiments, R¹ is —OR²⁰.

In some embodiments, R² is H. In some embodiments, R² is F. In someembodiments, R² is Cl. In some embodiments, R² is C₁-C₃ alkyl. In someembodiments, R² is —OR²⁰.

In some embodiments, R³ is H. In some embodiments, R³ is F. In someembodiments, R³ is Cl. In some embodiments, R³ is C₁-C₃ alkyl. In someembodiments, R³ is —OR²⁰.

In some embodiments, R¹ and R² together with the atoms they are attachedto form an optionally substituted 5-7 membered ring. In someembodiments, R² and R³ together with the atoms they are attached to forman optionally substituted 5-7 membered ring.

In some embodiments, R²⁰ is CH₂-R²¹. In some embodiments, R²⁰ is amethyl optionally substituted with 2 or 3 fluorine atoms. In someembodiments, R²⁰ is C₃-C₆ cycloalkyl. In some embodiments, R²⁰ is C₁-C₆alkyl.

In some embodiments, R²¹ is C₁-C₁₀ alkyl. In some embodiments, R²¹ is abranched C₃-C₁₀ alkyl optionally substituted with one or more hydroxy orfluoro. In some embodiments, R²¹ is C₃-C₆ cycloalkyl.

In some embodiments, R²¹ is

In some embodiments, R²² is an optionally substituted C₁-C₃ alkyl. Insome embodiments, R²² is hydroxy.

In some embodiments, R²³ is an optionally substituted C₁-C₃ alkyl. Insome embodiments, R²³ is hydroxy.

In some embodiments, R²⁴ is an optionally substituted C₁-C₃ alkyl. Insome embodiments, R²⁴ is hydroxy.

In some embodiments, two of R²²-R²⁴ together with the atoms they areattached to form an optionally substituted 5-7 membered ring.

In some embodiments, B is selected from the group consisting of:

In some embodiments, the alkoxy group is further substituted wherein1-5, preferably, 1-3 hydrogen atoms are substituted, preferredsubstituents including without limitation, C₁-C₆ alkyl optionallysubstituted with 1-3 halo, such as fluoro, and/or C₁-C₆ alkoxy;optionally substituted C₁-C₆ alkoxy; and halo, preferably fluoro. Insome embodiments, substituents include without limitation C₁-C₆ alkylsubstituted with 1-3 halo, such as fluoro. In some embodiments,substituents include without limitation C₁-C₆ alkyl optionallysubstituted with C₁-C₆ alkoxy. In some embodiments, substituents includewithout limitation a substituted C₁-C₆ alkoxy. In some embodiments,substituents include without limitation one or more halo. In someembodiments, substituents include one or more fluoro. In someembodiments, the ring moiety such as the cyclopropyl group is furthersubstituted with 1-3 halo, preferably 1-2 halo. In some embodiments, thering moiety, such as the cyclopropyl group, is further substituted with1-2 halo. In some embodiments, the methylene group between the oxygenatom and the ring moiety, such as the cyclopropyl group, is substitutedwith 1-2 C₁-C₆ alkyl, preferably methyl, ethyl, or propyl groups. Insome embodiments, the methylene group is substituted with methyl groups.In some embodiments, the methylene group is substituted with ethylgroups. In some embodiments, the methylene group is substituted withpropyl groups. In some embodiments, R⁷⁰ is a hydrogen or an optionallysubstituted C₁-C₁₀ alkyl.

In some embodiments, the alkoxy group is further optionally substitutedwherein 1-5 hydrogen atoms are optionally substituted. In someembodiments, substituents include without limitation C₁-C₆ alkyloptionally substituted with 1-3 halo, such as fluoro. In someembodiments, substituents include without limitation C₁-C₆ alkyloptionally substituted with C₁-C₆ alkoxy. In some embodiments,substituents include without limitation an optionally substituted C₁-C₆alkoxy. In some embodiments, substituents include without limitation ahalo. In some embodiments, substituents include a fluoro.

In some embodiments, the ring moiety such as the cyclopropyl group isfurther optionally substituted with 1-3 halo. In some embodiments, thering moiety, such as the cyclopropyl group, is further optionallysubstituted with 1-2 halo.

In some embodiments, the methylene group between the oxygen atom and thering moiety, such as the cyclopropyl group, is optionally substitutedwith 1-2 C₁-C₆ alkyl. In some embodiments, the methylene group isoptionally substituted with methyl groups. In some embodiments, themethylene group is optionally substituted with ethyl groups. In someembodiments, the methylene group is optionally substituted with propylgroups.

In some embodiments, B is:

In some embodiments, the compound of formula (I) is not

This disclosure also provides a tautomer, or its pharmaceuticallyacceptable salt of a compound as disclosed herein.

This disclosure also provides a stereochemically pure enantiomer of acompound as described herein, its tautomer, diastereoisomer or itspharmaceutically acceptable salt. Methods to purify and identify thepure enantiomer are known in the art and described herein.

In another aspect, compositions comprising one or more of theabove-noted compounds and a carrier are provided herein. In oneembodiment, the composition is a pharmaceutical composition andtherefore further comprise at least a pharmaceutically acceptablecarrier or a pharmaceutically acceptable excipient. The compositions areformulated for various delivery modes, e.g., systemic (oral) or local.

In another aspect, this disclosure provides compositions comprising oneor more compounds as provided herein and a dUTPase-directed chemotherapyand a carrier, such as a pharmaceutically acceptable carrier. Thecompound and chemotherapy can be in varying amounts, and in one aspect,each in an effective amount when used in combination, provides atherapeutic benefit as described herein. The compositions are formulatedfor various delivery modes, e.g., systemic (oral) or local.

In one aspect, provided is a composition comprising a compound providedherein and at least one pharmaceutically acceptable excipient orcarrier.

In another aspect, methods are provided for inhibiting deoxyuridinetriphosphatase (dUTPase) comprising contacting the dUTPase with atherapeutically effective amount of a compound or a composition providedherein. In another aspect, the method further comprises contacting thedUTPase with a dUTPase-directed chemotherapy alone or in combinationwith the compound as provided herein. The contacting can be in vitro, invivo, simultaneous or concurrent. In a further aspect thedUTPase-directed chemotherapy is contacted prior to the compound orcomposition as described herein. In another aspect, the dUTPase-directedchemotherapy is contacted subsequent to the compound or composition. Ina yet further aspect, the compound or composition and thedUTPase-directed chemotherapy are sequentially administered throughseveral rounds of therapy. The contacting can be simultaneous orconcurrent and/or in vitro (cell free), ex vivo or in vivo. In a furtheraspect, the compounds or compositions of this disclosure areadministered to a patient identified or selected for the therapy bydetermining that the patient has a tumor or mass that over expressesdUTPase. Methods to identify such patients are known in the art andincorporated herein. The methods when administered to a subject such asa human patient, can be first line, second line, third line, fourth lineor further therapy.

Also provided is a method for reversing resistance to a dUTPase-directedchemotherapy comprising contacting a cell overexpressing dUTPase with atherapeutically effective amount of a compound or a composition providedherein, alone or in combination with a dUTPase-directed chemotherapy. Inone aspect, the cell is first identified as overexpressing dUTPase by ascreen as disclosed by U.S. Pat. No. 5,962,246. In another aspect, themethod further comprises subsequently contacting the cell expressingdUTPase with a dUTPase-directed chemotherapy. The methods can beadministered as second line, third line, fourth line or further therapy.

Further provided is a method for enhancing the efficacy of adUTPase-directed chemotherapy comprising contacting a cell, e.g., in oneaspect a cell over expressing dUTPase, with a therapeutically effectiveamount of a compound or a composition provided herein. In anotheraspect, the method further comprises contacting the cell with adUTPase-directed chemotherapy. The contacting can be simultaneous orconcurrent and/or in vitro (cell free), ex vivo or in vivo. In a furtheraspect, the dUTPase-directed chemotherapy is contacted prior to thecompound or composition as described herein, or vice versa. The methodswhen administered to a subject such as a human patient, can be firstline, second line, third line, fourth line or further therapy.

In another aspect, provided herein is a method of treating a diseaseassociated with the dUTPase pathway, e.g., cancer, viral infection,bacterial infection, or an autoimmune disorder, comprising administeringto a patient in need of such treatment a therapeutically effectiveamount of the compound provided herein or a composition provided hereinin combination with an agent which is suitable for treating the disease,thereby treating the disease. The administration of the compound of thisinvention and the agent that is suitable for the disease (e.g., adUTPase inhibitor) can be simultaneous or concurrent and/or in vitro(cell free), ex vivo or in vivo. In a further aspect the agent that issuitable for treating the disease is administered prior to the compoundor composition as described herein, or vice versa. In one aspect, thepatient being treated is selected for the therapy by screening a cell ortissue sample isolated from the patient for over expression of dUTPase.The therapy is then administered to this patient after the screen, andthe patient has been selected for therapy.

In another aspect, provided herein is a method of inhibiting the growthof a cancer cell comprising contacting the cell with a therapeuticallyeffective amount of the compounds or compositions as disclosed hereinand an effective amount of a dUTPase directed therapeutic, therebyinhibiting the growth of the cancer cell.

In another aspect, provided herein is a kit comprising a compoundprovided herein or a composition provided herein. The kit can furthercomprise one more of a dUTPase inhibitor (e.g., an antitumor agent) andinstructions for administering the agent. Yet further provided in thekit are reagents and instructions to screen for dUTPase expression.

In each of the above embodiments, a non-limiting example of the dUTPasemediated chemotherapy comprises a TS-inhibitor, e.g., 5-FU or 5-FUcontaining therapy such as 5-FU based adjuvant therapy and chemicalequivalents thereof.

In one aspect, provided is a method of one or more of inhibiting dUTPaseor enhancing the efficacy of a dUTPase directed therapy comprisingcontacting the dUTPase with a therapeutically effective amount of thecompound or composition provided herein.

In one aspect, provided is a method of reversing resistance to adUTPase-directed therapy comprising contacting the dUTPase with atherapeutically effective amount of the compound or composition providedherein.

In one aspect, provided is a method of treating a disease whosetreatment is impeded by the expression or over expression of dUTPase,comprising administering to a patient in need of such treatment atherapeutically effective amount of the compound or composition providedherein.

In one aspect, provided is a method of inhibiting the growth of a cancercell comprising contacting the cell with a therapeutically effectiveamount of the compound or composition provided herein and atherapeutically effective amount of a dUTPase directed therapeutic,thereby inhibiting the growth of the cancer cell.

In some embodiments, the cancer cell is selected from a colon cancercell, a colorectal cancer cell, a gastric cancer cell, a head and neckcancer cell, a breast cancer cell, a lung cancer cell or a blood cell.

In one aspect, provided is a method of treating a disease in a patientwhose treatment is impeded by the expression or overexpression ofdUTPase, comprising: a) screening a cell or tissue sample from thepatient; b) determining the expression level of dUTPase in the sample;and c) administering to a patient whose sample shows over expression ofdUTPase, a therapeutically effective amount of the compound orcomposition provided herein.

In some embodiments, the disease is cancer. In some embodiments, thecancer is selected from the group consisting of colon cancer, colorectalcancer, gastric cancer, esophageal cancer, head and neck cancer, breastcancer, lung cancer, stomach cancer, liver cancer, gall bladder cancer,or pancreatic cancer or leukemia.

In one aspect, provided is a kit comprising a compound or compositionprovided herein and instructions for use in a diagnostic or therapeuticmethod as described herein.

DETAILED DESCRIPTION Definitions

Throughout this disclosure, various publications, patents and publishedpatent specifications are referenced by an identifying citation. Thedisclosures of these publications, patents and published patentspecifications are hereby incorporated by reference into the presentdisclosure in their entirety to more fully describe the state of the artto which this invention pertains.

The practice of the present technology will employ, unless otherwiseindicated, conventional techniques of organic chemistry, pharmacology,immunology, molecular biology, microbiology, cell biology andrecombinant DNA, which are within the skill of the art. See, e.g.,Sambrook, Fritsch and Maniatis, Molecular Cloning: A Laboratory Manual,2^(nd) edition (1989); Current Protocols In Molecular Biology (F. M.Ausubel, et al. eds., (1987)); the series Methods in Enzymology(Academic Press, Inc.): PCR 2: A Practical Approach (M. J. MacPherson,B. D. Hames and G. R. Taylor eds. (1995)), Harlow and Lane, eds. (1988)Antibodies, a Laboratory Manual, and Animal Cell Culture (R. I.Freshney, ed. (1987)).

As used in the specification and claims, the singular form “a,” “an” and“the” include plural references unless the context clearly dictatesotherwise. For example, the term “a cell” includes a plurality of cells,including mixtures thereof.

As used herein, the term “comprising” is intended to mean that thecompounds, compositions and methods include the recited elements, butnot exclude others. “Consisting essentially of” when used to definecompounds, compositions and methods, shall mean excluding other elementsof any essential significance to the combination. Thus, a compositionconsisting essentially of the elements as defined herein would notexclude trace contaminants, e.g., from the isolation and purificationmethod and pharmaceutically acceptable carriers, preservatives, and thelike. “Consisting of” shall mean excluding more than trace elements ofother ingredients. Embodiments defined by each of these transition termsare within the scope of this technology.

All numerical designations, e.g., pH, temperature, time, concentration,and molecular weight, including ranges, are approximations which arevaried (+) or (−) by increments of 1, 5, or 10%. It is to be understood,although not always explicitly stated that all numerical designationsare preceded by the term “about.” It also is to be understood, althoughnot always explicitly stated, that the reagents described herein aremerely exemplary and that equivalents of such are known in the art.

“Alkyl” refers to monovalent saturated aliphatic hydrocarbyl groupshaving from 1 to 10 carbon atoms and preferably 1 to 6 carbon atoms.This term includes, by way of example, linear and branched hydrocarbylgroups such as methyl (CH₃—), ethyl (CH₃CH₂—), n-propyl (CH₃CH₂CH₂—),isopropyl ((CH₃)₂CH—), n-butyl (CH₃CH₂CH₂CH₂—), isobutyl ((CH₃)₂CHCH₂—),sec-butyl ((CH₃)(CH₃CH₂)CH—), t-butyl ((CH₃)₃C—), n-pentyl(CH₃CH₂CH₂CH₂CH₂—), and neopentyl ((CH₃)₃CCH₂—).

“Alkenyl” refers to monovalent straight or branched hydrocarbyl groupshaving from 2 to 10 carbon atoms and preferably 2 to 6 carbon atoms orpreferably 2 to 4 carbon atoms and having at least 1 and preferably from1 to 2 sites of vinyl (>C═C<) unsaturation. Such groups are exemplified,for example, by vinyl, allyl, and but-3-en-1-yl. Included within thisterm are the cis and trans isomers or mixtures of these isomers.

“Alkynyl” refers to straight or branched monovalent hydrocarbyl groupshaving from 2 to 10 carbon atoms and preferably 2 to 6 carbon atoms orpreferably 2 to 3 carbon atoms and having at least 1 and preferably from1 to 2 sites of acetylenic (—C≡C—) unsaturation. Examples of suchalkynyl groups include acetylenyl (—C≡CH), and propargyl (—CH₂C≡CH).

“Substituted alkyl” refers to an alkyl group having from 1 to 5,preferably 1 to 3, or more preferably 1 to 2 substituents selected fromthe group consisting of alkoxy, substituted alkoxy, acyl, acylamino,acyloxy, amino, substituted amino, aminocarbonyl, aminothiocarbonyl,aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy,aminosulfonyl, aminosulfonyloxy, aminosulfonylamino, amidino, aryl,substituted aryl, aryloxy, substituted aryloxy, arylthio, substitutedarylthio, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxylester)oxy, cyano, cycloalkyl, substituted cycloalkyl, cycloalkyloxy,substituted cycloalkyloxy, cycloalkylthio, substituted cycloalkylthio,cycloalkenyl, substituted cycloalkenyl, cycloalkenyloxy, substitutedcycloalkenyloxy, cycloalkenylthio, substituted cycloalkenylthio,guanidino, substituted guanidino, halo, hydroxy, heteroaryl, substitutedheteroaryl, heteroaryloxy, substituted heteroaryloxy, heteroarylthio,substituted heteroarylthio, heterocyclic, substituted heterocyclic,heterocyclyloxy, substituted heterocyclyloxy, heterocyclylthio,substituted heterocyclylthio, nitro, SO₃H, substituted sulfonyl,substituted sulfonyloxy, thioacyl, thiol, alkylthio, and substitutedalkylthio, wherein said substituents are as defined herein.

“Heteroalkyl” refers to an alkyl group one or more carbons is replacedwith —O—, —S—, SO₂, a P containing moiety as provided herein, —NR^(Q)—,

moieties where R^(Q) is H or C₁-C₆ alkyl. Substituted heteroalkyl refersto a heteroalkyl group having from 1 to 5, preferably 1 to 3, or morepreferably 1 to 2 substituents selected from the group consisting ofalkoxy, substituted alkoxy, acyl, acylamino, acyloxy, amino, substitutedamino, aminocarbonyl, aminothiocarbonyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, aminosulfonyl,aminosulfonyloxy, aminosulfonylamino, amidino, aryl, substituted aryl,aryloxy, substituted aryloxy, arylthio, substituted arylthio, carboxyl,carboxyl ester, (carboxyl ester)amino, (carboxyl ester)oxy, cyano,cycloalkyl, substituted cycloalkyl, cycloalkyloxy, substitutedcycloalkyloxy, cycloalkylthio, substituted cycloalkylthio, cycloalkenyl,substituted cycloalkenyl, cycloalkenyloxy, substituted cycloalkenyloxy,cycloalkenylthio, substituted cycloalkenylthio, guanidino, substitutedguanidino, halo, hydroxy, heteroaryl, substituted heteroaryl,heteroaryloxy, substituted heteroaryloxy, heteroarylthio, substitutedheteroarylthio, heterocyclic, substituted heterocyclic, heterocyclyloxy,substituted heterocyclyloxy, heterocyclylthio, substitutedheterocyclylthio, nitro, SO₃H, substituted sulfonyl, substitutedsulfonyloxy, thioacyl, thiol, alkylthio, and substituted alkylthio,wherein said substituents are as defined herein.

“Substituted alkenyl” refers to alkenyl groups having from 1 to 3substituents, and preferably 1 to 2 substituents, selected from thegroup consisting of alkoxy, substituted alkoxy, acyl, acylamino,acyloxy, amino, substituted amino, aminocarbonyl, aminothiocarbonyl,aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy,aminosulfonyl, aminosulfonyloxy, aminosulfonylamino, amidino, aryl,substituted aryl, aryloxy, substituted aryloxy, arylthio, substitutedarylthio, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxylester)oxy, cyano, cycloalkyl, substituted cycloalkyl, cycloalkyloxy,substituted cycloalkyloxy, cycloalkylthio, substituted cycloalkylthio,cycloalkenyl, substituted cycloalkenyl, cycloalkenyloxy, substitutedcycloalkenyloxy, cycloalkenylthio, substituted cycloalkenylthio,guanidino, substituted guanidino, halo, hydroxyl, heteroaryl,substituted heteroaryl, heteroaryloxy, substituted heteroaryloxy,heteroarylthio, substituted heteroarylthio, heterocyclic, substitutedheterocyclic, heterocyclyloxy, substituted heterocyclyloxy,heterocyclylthio, substituted heterocyclylthio, nitro, SO₃H, substitutedsulfonyl, substituted sulfonyloxy, thioacyl, thiol, alkylthio, andsubstituted alkylthio, wherein said substituents are as defined hereinand with the proviso that any hydroxyl or thiol substitution is notattached to a vinyl (unsaturated) carbon atom.

“Heteroalkenyl” refers to an alkenyl group one or more carbons isreplaced with —O—, —S—, SO₂, a P containing moiety as provided herein,—NR^(Q)—,

moieties where R^(Q) is H or C₁-C₆ alkyl. Substituted heteroalkenylrefers to a heteroalkenyl group having from 1 to 5, preferably 1 to 3,or more preferably 1 to 2 substituents selected from the groupconsisting of alkoxy, substituted alkoxy, acyl, acylamino, acyloxy,amino, substituted amino, aminocarbonyl, aminothiocarbonyl,aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy,aminosulfonyl, aminosulfonyloxy, aminosulfonylamino, amidino, aryl,substituted aryl, aryloxy, substituted aryloxy, arylthio, substitutedarylthio, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxylester)oxy, cyano, cycloalkyl, substituted cycloalkyl, cycloalkyloxy,substituted cycloalkyloxy, cycloalkylthio, substituted cycloalkylthio,cycloalkenyl, substituted cycloalkenyl, cycloalkenyloxy, substitutedcycloalkenyloxy, cycloalkenylthio, substituted cycloalkenylthio,guanidino, substituted guanidino, halo, hydroxy, heteroaryl, substitutedheteroaryl, heteroaryloxy, substituted heteroaryloxy, heteroarylthio,substituted heteroarylthio, heterocyclic, substituted heterocyclic,heterocyclyloxy, substituted heterocyclyloxy, heterocyclylthio,substituted heterocyclylthio, nitro, SO₃H, substituted sulfonyl,substituted sulfonyloxy, thioacyl, thiol, alkylthio, and substitutedalkylthio, wherein said substituents are as defined herein.

“Substituted alkynyl” refers to alkynyl groups having from 1 to 3substituents, and preferably 1 to 2 substituents, selected from thegroup consisting of alkoxy, substituted alkoxy, acyl, acylamino,acyloxy, amino, substituted amino, aminocarbonyl, aminothiocarbonyl,aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy,aminosulfonyl, aminosulfonyloxy, aminosulfonylamino, amidino, aryl,substituted aryl, aryloxy, substituted aryloxy, arylthio, substitutedarylthio, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxylester)oxy, cyano, cycloalkyl, substituted cycloalkyl, cycloalkyloxy,substituted cycloalkyloxy, cycloalkylthio, substituted cycloalkylthio,cycloalkenyl, substituted cycloalkenyl, cycloalkenyloxy, substitutedcycloalkenyloxy, cycloalkenylthio, substituted cycloalkenylthio,guanidino, substituted guanidino, halo, hydroxy, heteroaryl, substitutedheteroaryl, heteroaryloxy, substituted heteroaryloxy, heteroarylthio,substituted heteroarylthio, heterocyclic, substituted heterocyclic,heterocyclyloxy, substituted heterocyclyloxy, heterocyclylthio,substituted heterocyclylthio, nitro, SO₃H, substituted sulfonyl,substituted sulfonyloxy, thioacyl, thiol, alkylthio, and substitutedalkylthio, wherein said substituents are as defined herein and with theproviso that any hydroxyl or thiol substitution is not attached to anacetylenic carbon atom.

“Heteroalkynyl” refers to an alkynyl group one or more carbons isreplaced with —O—, —S—, SO₂, a P containing moiety as provided herein,—NR^(Q)—,

moieties where R^(Q) is H or C₁-C₆ alkyl. Substituted heteroalkynylrefers to a heteroalkynyl group having from 1 to 5, preferably 1 to 3,or more preferably 1 to 2 substituents selected from the groupconsisting of alkoxy, substituted alkoxy, acyl, acylamino, acyloxy,amino, substituted amino, aminocarbonyl, aminothiocarbonyl,aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy,aminosulfonyl, aminosulfonyloxy, aminosulfonylamino, amidino, aryl,substituted aryl, aryloxy, substituted aryloxy, arylthio, substitutedarylthio, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxylester)oxy, cyano, cycloalkyl, substituted cycloalkyl, cycloalkyloxy,substituted cycloalkyloxy, cycloalkylthio, substituted cycloalkylthio,cycloalkenyl, substituted cycloalkenyl, cycloalkenyloxy, substitutedcycloalkenyloxy, cycloalkenylthio, substituted cycloalkenylthio,guanidino, substituted guanidino, halo, hydroxy, heteroaryl, substitutedheteroaryl, heteroaryloxy, substituted heteroaryloxy, heteroarylthio,substituted heteroarylthio, heterocyclic, substituted heterocyclic,heterocyclyloxy, substituted heterocyclyloxy, heterocyclylthio,substituted heterocyclylthio, nitro, SO₃H, substituted sulfonyl,substituted sulfonyloxy, thioacyl, thiol, alkylthio, and substitutedalkylthio, wherein said substituents are as defined herein.

“Alkylene” refers to divalent saturated aliphatic hydrocarbyl groupshaving from 1 to 10 carbon atoms, preferably having from 1 to 6 and morepreferably 1 to 3 carbon atoms that are either straight-chained orbranched. This term is exemplified by groups such as methylene (—CH₂—),ethylene (—CH₂CH₂—), n-propylene (—CH₂CH₂CH₂—), iso-propylene(—CH₂CH(CH₃)— or —CH(CH₃)CH₂—), butylene (—CH₂CH₂CH₂CH₂—), isobutylene(—CH₂CH(CH₃)CH₂—), sec-butylene (—CH₂CH₂(CH₃)CH—), and the like.Similarly, “alkenylene” and “alkynylene” refer to an alkylene moietycontaining respective 1 or 2 carbon carbon double bonds or a carboncarbon triple bond.

“Substituted alkylene” refers to an alkylene group having from 1 to 3hydrogens replaced with substituents selected from the group consistingof alkyl, substituted alkyl, alkoxy, substituted alkoxy, acyl,acylamino, acyloxy, amino, substituted amino, aminoacyl, aryl,substituted aryl, aryloxy, substituted aryloxy, cyano, halogen,hydroxyl, nitro, carboxyl, carboxyl ester, cycloalkyl, substitutedcycloalkyl, heteroaryl, substituted heteroaryl, heterocyclic,substituted heterocyclic, and oxo wherein said substituents are definedherein. In some embodiments, the alkylene has 1 to 2 of theaforementioned groups, or having from 1-3 carbon atoms replaced with—O—, —S—, or —NR^(Q)— moieties where R^(Q) is H or C₁-C₆ alkyl. It is tobe noted that when the alkylene is substituted by an oxo group, 2hydrogens attached to the same carbon of the alkylene group are replacedby “═O”. “Substituted alkenylene” and “substituted alkynylene” refer toalkenylene and substituted alkynylene moieties substituted withsubstituents as described for substituted alkylene.

“Alkynylene” refers to straight or branched divalent hydrocarbyl groupshaving from 2 to 10 carbon atoms and preferably 2 to 6 carbon atoms orpreferably 2 to 3 carbon atoms and having at least 1 and preferably from1 to 2 sites of acetylenic (—C≡C—) unsaturation. Examples of suchalkynylene groups include —C≡C— and —CH₂C≡C—.

“Substituted alkynylene” refers to alkynylene groups having from 1 to 3substituents, and preferably 1 to 2 substituents, selected from thegroup consisting of alkoxy, substituted alkoxy, acyl, acylamino,acyloxy, amino, substituted amino, aminocarbonyl, aminothiocarbonyl,aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy,aminosulfonyl, aminosulfonyloxy, aminosulfonylamino, amidino, aryl,substituted aryl, aryloxy, substituted aryloxy, arylthio, substitutedarylthio, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxylester)oxy, cyano, cycloalkyl, substituted cycloalkyl, cycloalkyloxy,substituted cycloalkyloxy, cycloalkylthio, substituted cycloalkylthio,cycloalkenyl, substituted cycloalkenyl, cycloalkenyloxy, substitutedcycloalkenyloxy, cycloalkenylthio, substituted cycloalkenylthio,guanidino, substituted guanidino, halo, hydroxy, heteroaryl, substitutedheteroaryl, heteroaryloxy, substituted heteroaryloxy, heteroarylthio,substituted heteroarylthio, heterocyclic, substituted heterocyclic,heterocyclyloxy, substituted heterocyclyloxy, heterocyclylthio,substituted heterocyclylthio, nitro, SO₃H, substituted sulfonyl,substituted sulfonyloxy, thioacyl, thiol, alkylthio, and substitutedalkylthio, wherein said substituents are as defined herein and with theproviso that any hydroxyl or thiol substitution is not attached to anacetylenic carbon atom.

“Heteroalkylene” refers to an alkylene group wherein one or more carbonsis replaced with —O—, —S—, SO₂, a P containing moiety as providedherein, —NR^(Q)—,

moieties where R^(Q) is H or C₁-C₆ alkyl. “Substituted heteroalkylene”refers to heteroalkynylene groups having from 1 to 3 substituents, andpreferably 1 to 2 substituents, selected from the substituents disclosedfor substituted alkylene.

“Heteroalkenylene” refers to an alkenylene group wherein one or morecarbons is replaced with —O—, —S—, SO₂, a P containing moiety asprovided herein, —NR^(Q)—,

moieties where R^(Q) is H or C₁-C₆ alkyl. “Substituted heteroalkenylene”refers to heteroalkynylene groups having from 1 to 3 substituents, andpreferably 1 to 2 substituents, selected from the substituents disclosedfor substituted alkenylene.

“Heteroalkynylene” refers to an alkynylene group wherein one or morecarbons is replaced with —O—, —S—, SO₂, a P containing moiety asprovided herein, —NR^(Q)—,

moieties where R^(Q) is H or C₁-C₆ alkyl. “Substituted heteroalkynylene”refers to heteroalkynylene groups having from 1 to 3 substituents, andpreferably 1 to 2 substituents, selected from the substituents disclosedfor substituted alkynylene.

“Alkoxy” refers to the group —O-alkyl wherein alkyl is defined herein.Alkoxy includes, by way of example, methoxy, ethoxy, n-propoxy,isopropoxy, n-butoxy, t-butoxy, sec-butoxy, and n-pentoxy.

“Substituted alkoxy” refers to the group —O-(substituted alkyl) whereinsubstituted alkyl is defined herein.

“Acyl” refers to the groups H—C(O)—, alkyl-C(O)—, substitutedalkyl-C(O)—, alkenyl-C(O)—, substituted alkenyl-C(O)—, alkynyl-C(O)—,substituted alkynyl-C(O)—, cycloalkyl-C(O)—, substitutedcycloalkyl-C(O)—, cycloalkenyl-C(O)—, substituted cycloalkenyl-C(O)—,aryl-C(O)—, substituted aryl-C(O)—, heteroaryl-C(O)—, substitutedheteroaryl-C(O)—, heterocyclic-C(O)—, and substitutedheterocyclic-C(O)—, wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic are as defined herein. Acyl includes the“acetyl” group CH₃C(O)—.

“Acylamino” refers to the groups —NR⁴⁷C(O)alkyl, —NR⁴⁷C(O)substitutedalkyl, —NR⁴⁷C(O)cycloalkyl, —NR⁴⁷C(O)substituted cycloalkyl,—NR⁴⁷C(O)cycloalkenyl, —NR⁴⁷C(O)substituted cycloalkenyl,—NR⁴⁷C(O)alkenyl, —NR⁴⁷C(O)substituted alkenyl, —NR⁴⁷C(O)alkynyl,—NR⁴⁷C(O)substituted alkynyl, —NR⁴⁷C(O)aryl, —NR⁴⁷C(O)substituted aryl,—NR⁴⁷C(O)heteroaryl, —NR⁴⁷C(O)substituted heteroaryl,—NR⁴⁷C(O)heterocyclic, and —NR⁴⁷C(O)substituted heterocyclic wherein R⁴⁷is hydrogen or alkyl and wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic are as defined herein.

“Acyloxy” refers to the groups alkyl-C(O)O—, substituted alkyl-C(O)O—,alkenyl-C(O)O—, substituted alkenyl-C(O)O—, alkynyl-C(O)O—, substitutedalkynyl-C(O)O—, aryl-C(O)O—, substituted aryl-C(O)O—, cycloalkyl-C(O)O—,substituted cycloalkyl-C(O)O—, cycloalkenyl-C(O)O—, substitutedcycloalkenyl-C(O)O—, heteroaryl-C(O)O—, substituted heteroaryl-C(O)O—,heterocyclic-C(O)O—, and substituted heterocyclic-C(O)O— wherein alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, and substituted heterocyclic are as definedherein.

An animal, subject or patient for diagnosis or treatment refers to ananimal such as a mammal, or a human, ovine, bovine, feline, canine,equine, simian, etc. Non-human animals subject to diagnosis or treatmentinclude, for example, simians, murine, such as, rat, mice, canine,leporid, livestock, sport animals, and pets.

“Amino” refers to the group —NH₂.

“Substituted amino” refers to the group —NR⁴⁸R⁴⁹ where R⁴⁸ and R⁴⁹ areindependently selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-alkenyl, —SO₂-substituted alkenyl,—SO₂-cycloalkyl, —SO₂-substituted cylcoalkyl, —SO₂-cycloalkenyl,—SO₂-substituted cylcoalkenyl, —SO₂-aryl, —SO₂-substituted aryl,—SO₂-heteroaryl, —SO₂-substituted heteroaryl, —SO₂-heterocyclic, and—SO₂-substituted heterocyclic and wherein R⁴⁸ and R⁴⁹ are optionallyjoined, together with the nitrogen bound thereto to form a heterocyclicor substituted heterocyclic group, provided that R⁴⁸ and R⁴⁹ are bothnot hydrogen, and wherein alkyl, substituted alkyl, alkenyl, substitutedalkenyl, alkynyl, substituted alkynyl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, heterocyclic, and substitutedheterocyclic are as defined herein. When R⁴⁸ is hydrogen and R⁴⁹ isalkyl, the substituted amino group is sometimes referred to herein asalkylamino. When R⁴⁸ and R⁴⁹ are alkyl, the substituted amino group issometimes referred to herein as dialkylamino. When referring to amonosubstituted amino, it is meant that either R⁴⁸ or R⁴⁹ is hydrogenbut not both. When referring to a disubstituted amino, it is meant thatneither R⁴⁸ nor R⁴⁹ are hydrogen.

“Aminocarbonyl” refers to the group —C(O)NR⁵⁰R⁵¹ where R⁵⁰ and R⁵¹ areindependently selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, heteroaryl, substitutedheteroaryl, heterocyclic, and substituted heterocyclic and where R⁵⁰ andR⁵¹ are optionally joined together with the nitrogen bound thereto toform a heterocyclic or substituted heterocyclic group, and whereinalkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, and substituted heterocyclic areas defined herein.

“Aminothiocarbonyl” refers to the group —C(S)NR⁵⁰R⁵¹ where R⁵⁰ and R⁵¹are independently selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, heteroaryl, substitutedheteroaryl, heterocyclic, and substituted heterocyclic and where R⁵⁰ andR⁵¹ are optionally joined together with the nitrogen bound thereto toform a heterocyclic or substituted heterocyclic group, and whereinalkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, and substituted heterocyclic areas defined herein.

“Aminocarbonylamino” refers to the group —NR⁴⁷C(O)NR⁵⁰R⁵¹ where R⁴⁷ ishydrogen or alkyl and R⁵⁰ and R⁵¹ are independently selected from thegroup consisting of hydrogen, alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, aryl, substitutedaryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic, and where R⁵⁰ and R⁵¹ are optionally joinedtogether with the nitrogen bound thereto to form a heterocyclic orsubstituted heterocyclic group, and wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic are as defined herein.

“Aminothiocarbonylamino” refers to the group —NR⁴⁷C(S)NR⁵⁰R⁵¹ where R⁴⁷is hydrogen or alkyl and R⁵⁰ and R⁵¹ are independently selected from thegroup consisting of hydrogen, alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, aryl, substitutedaryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic and where R⁵⁰ and R⁵¹ are optionally joinedtogether with the nitrogen bound thereto to form a heterocyclic orsubstituted heterocyclic group, and wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic are as defined herein.

“Aminocarbonyloxy” refers to the group —O—C(O)NR⁵⁰R⁵¹ where R⁵⁰ and R⁵¹are independently selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, heteroaryl, substitutedheteroaryl, heterocyclic, and substituted heterocyclic and where R⁵⁰ andR⁵¹ are optionally joined together with the nitrogen bound thereto toform a heterocyclic or substituted heterocyclic group, and whereinalkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, and substituted heterocyclic areas defined herein.

“Aminosulfonyl” refers to the group —SO₂NR⁵⁰R⁵¹ where R⁵⁰ and R⁵¹ areindependently selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, heteroaryl, substitutedheteroaryl, heterocyclic, and substituted heterocyclic and where R⁵⁰ andR⁵¹ are optionally joined together with the nitrogen bound thereto toform a heterocyclic or substituted heterocyclic group, and whereinalkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, and substituted heterocyclic areas defined herein.

“Aminosulfonyloxy” refers to the group —O—SO₂NR⁵⁰R⁵¹ where R⁵⁰ and R⁵¹are independently selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, heteroaryl, substitutedheteroaryl, heterocyclic, and substituted heterocyclic and where R⁵⁰ andR⁵¹ are optionally joined together with the nitrogen bound thereto toform a heterocyclic or substituted heterocyclic group, and whereinalkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, and substituted heterocyclic areas defined herein.

“Aminosulfonylamino” refers to the group —NR⁴⁷SO₂NR⁵⁰R⁵¹ where R⁴⁷ ishydrogen or alkyl and R⁵⁰ and R⁵¹ are independently selected from thegroup consisting of hydrogen, alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, aryl, substitutedaryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic and where R⁵⁰ and R⁵¹ are optionally joinedtogether with the nitrogen bound thereto to form a heterocyclic orsubstituted heterocyclic group, and wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic are as defined herein.

“Amidino” refers to the group —C(═NR⁵²)NR⁵⁰R⁵¹ where R⁵⁰, R⁵¹, and R⁵²are independently selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, heteroaryl, substitutedheteroaryl, heterocyclic, and substituted heterocyclic and where R⁵⁰ andR⁵¹ are optionally joined together with the nitrogen bound thereto toform a heterocyclic or substituted heterocyclic group, and whereinalkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, and substituted heterocyclic areas defined herein.

“Aryl” or “Ar” refers to a monovalent aromatic carbocyclic group of from6 to 14 carbon atoms having a single ring (e.g., phenyl) or multiplecondensed rings (e.g., naphthyl or anthryl) which condensed rings may ormay not be aromatic (e.g., 2-benzoxazolinone,2H-1,4-benzoxazin-3(4H)-one-7-yl, and the like) provided that the pointof attachment is at an aromatic carbon atom. Preferred aryl groupsinclude phenyl and naphthyl.

“Substituted aryl” refers to aryl groups which are substituted with 1 to5, preferably 1 to 3, or more preferably 1 to 2 substituents selectedfrom the group consisting of alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substitutedalkoxy, acyl, acylamino, acyloxy, amino, substituted amino,aminocarbonyl, aminothiocarbonyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, aminosulfonyl,aminosulfonyloxy, aminosulfonylamino, amidino, aryl, substituted aryl,aryloxy, substituted aryloxy, arylthio, substituted arylthio, carboxyl,carboxyl ester, (carboxyl ester)amino, (carboxyl ester)oxy, cyano,cycloalkyl, substituted cycloalkyl, cycloalkyloxy, substitutedcycloalkyloxy, cycloalkylthio, substituted cycloalkylthio, cycloalkenyl,substituted cycloalkenyl, cycloalkenyloxy, substituted cycloalkenyloxy,cycloalkenylthio, substituted cycloalkenylthio, guanidino, substitutedguanidino, halo, hydroxy, heteroaryl, substituted heteroaryl,heteroaryloxy, substituted heteroaryloxy, heteroarylthio, substitutedheteroarylthio, heterocyclic, substituted heterocyclic, heterocyclyloxy,substituted heterocyclyloxy, heterocyclylthio, substitutedheterocyclylthio, nitro, SO₃H, substituted sulfonyl, substitutedsulfonyloxy, thioacyl, thiol, alkylthio, and substituted alkylthio,wherein said substituents are as defined herein.

“Arylene” refers to a divalent aromatic carbocyclic group of from 6 to14 carbon atoms having a single ring or multiple condensed rings.“Substituted arylene” refers to an arylene having from 1 to 5,preferably 1 to 3, or more preferably 1 to 2 substituents as defined foraryl groups.

“Heteroarylene” refers to a divalent aromatic group of from 1 to 10carbon atoms and 1 to 4 heteroatoms selected from the group consistingof oxygen, nitrogen and sulfur within the ring. “Substitutedheteroarylene” refers to heteroarylene groups that are substituted withfrom 1 to 5, preferably 1 to 3, or more preferably 1 to 2 substituentsselected from the group consisting of the same group of substituentsdefined for substituted aryl.

“Aryloxy” refers to the group —O-aryl, where aryl is as defined herein,that includes, by way of example, phenoxy and naphthoxy.

“Substituted aryloxy” refers to the group —O-(substituted aryl) wheresubstituted aryl is as defined herein.

“Arylthio” refers to the group —S-aryl, where aryl is as defined herein.

“Substituted arylthio” refers to the group —S-(substituted aryl), wheresubstituted aryl is as defined herein.

“Carbonyl” refers to the divalent group —C(O)— which is equivalent to—C(═O)—.

“Carboxyl” or “carboxy” refers to —COOH or salts thereof.

“Carboxyl ester” or “carboxy ester” refers to the group —C(O)(O)-alkyl,—C(O)(O)-substituted alkyl, —C(O)O-alkenyl, —C(O)(O)-substitutedalkenyl, —C(O)(O)-alkynyl, —C(O)(O)-substituted alkynyl, —C(O)(O)-aryl,—C(O)(O)-substituted-aryl, —C(O)(O)-cycloalkyl, —C(O)(O)-substitutedcycloalkyl, —C(O)(O)-cycloalkenyl, —C(O)(O)-substituted cycloalkenyl,—C(O)(O)-heteroaryl, —C(O)(O)-substituted heteroaryl,—C(O)(O)-heterocyclic, and —C(O)(O)-substituted heterocyclic whereinalkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, and substituted heterocyclic areas defined herein.

“(Carboxyl ester)amino refers to the group —NR47C(O)(O)-alkyl,—NR47C(O)(O)-substituted alkyl, —NR47C(O)O-alkenyl,—NR47C(O)(O)-substituted alkenyl, —NR47C(O)(O)-alkynyl,—NR47C(O)(O)-substituted alkynyl, —NR47C(O)(O)-aryl,—NR47C(O)(O)-substituted-aryl, —NR47C(O)(O)-cycloalkyl,—NR47C(O)(O)-substituted cycloalkyl, —NR47C(O)(O)-cycloalkenyl,—NR47C(O)(O)-substituted cycloalkenyl, —NR47C(O)(O)-heteroaryl,—NR47C(O)(O)-substituted heteroaryl, —NR47C(O)(O)-heterocyclic, and—NR47C(O)(O)-substituted heterocyclic wherein R47 is alkyl or hydrogen,and wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl,alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocyclic, and substitutedheterocyclic are as defined herein.

“(Carboxyl ester)oxy refers to the group —O—C(O)O-alkyl,—O—C(O)O-substituted alkyl, —O—C(O)O-alkenyl, —O—C(O)O-substitutedalkenyl, —O—C(O)O-alkynyl, —O—C(O)(O)-substituted alkynyl,—O—C(O)O-aryl, —O—C(O)O-substituted-aryl, —O—C(O)O-cycloalkyl,—O—C(O)O-substituted cycloalkyl, —O—C(O)O-cycloalkenyl,—O—C(O)O-substituted cycloalkenyl, —O—C(O)O-heteroaryl,—O—C(O)O-substituted heteroaryl, —O—C(O)O-heterocyclic, and—O—C(O)O-substituted heterocyclic wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic are as defined herein.

A “composition” as used herein, refers to an active agent, such as acompound as disclosed herein and a carrier, inert or active. The carriercan be, without limitation, solid such as a bead or resin, or liquid,such as phosphate buffered saline.

Administration or treatment in “combination” refers to administering twoagents such that their pharmacological effects are manifest at the sametime. Combination does not require administration at the same time orsubstantially the same time, although combination can include suchadministrations.

“Cyano” refers to the group —CN.

“Cycloalkyl” refers to cyclic alkyl groups of from 3 to 10 carbon atomshaving single or multiple cyclic rings including fused, bridged, andspiro ring systems. The fused ring can be an aryl ring provided that thenon aryl part is joined to the rest of the molecule. Examples ofsuitable cycloalkyl groups include, for instance, adamantyl,cyclopropyl, cyclobutyl, cyclopentyl, and cyclooctyl.

“Cycloalkenyl” refers to non-aromatic cyclic alkyl groups of from 3 to10 carbon atoms having single or multiple cyclic rings and having atleast one >C═C< ring unsaturation and preferably from 1 to 2 sitesof >C═C< ring unsaturation.

“Substituted cycloalkyl” and “substituted cycloalkenyl” refers to acycloalkyl or cycloalkenyl group having from 1 to 5 or preferably 1 to 3substituents selected from the group consisting of oxo, thioxo, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, alkoxy, substituted alkoxy, acyl, acylamino, acyloxy, amino,substituted amino, aminocarbonyl, aminothiocarbonyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, aminosulfonyl,aminosulfonyloxy, aminosulfonylamino, amidino, aryl, substituted aryl,aryloxy, substituted aryloxy, arylthio, substituted arylthio, carboxyl,carboxyl ester, (carboxyl ester)amino, (carboxyl ester)oxy, cyano,cycloalkyl, substituted cycloalkyl, cycloalkyloxy, substitutedcycloalkyloxy, cycloalkylthio, substituted cycloalkylthio, cycloalkenyl,substituted cycloalkenyl, cycloalkenyloxy, substituted cycloalkenyloxy,cycloalkenylthio, substituted cycloalkenylthio, guanidino, substitutedguanidino, halo, hydroxy, heteroaryl, substituted heteroaryl,heteroaryloxy, substituted heteroaryloxy, heteroarylthio, substitutedheteroarylthio, heterocyclic, substituted heterocyclic, heterocyclyloxy,substituted heterocyclyloxy, heterocyclylthio, substitutedheterocyclylthio, nitro, SO₃H, substituted sulfonyl, substitutedsulfonyloxy, thioacyl, thiol, alkylthio, and substituted alkylthio,wherein said substituents are as defined herein.

“Cyclopropano” refers to:

“Cyclobutano” refers to:

“Cycloalkyloxy” refers to —O-cycloalkyl.

“Substituted cycloalkyloxy” refers to —O-(substituted cycloalkyl).

“Cycloalkylthio” refers to —S-cycloalkyl.

“Substituted cycloalkylthio” refers to —S-(substituted cycloalkyl).

“Cycloalkenyloxy” refers to —O-cycloalkenyl.

“Substituted cycloalkenyloxy” refers to —O-(substituted cycloalkenyl).

“Cycloalkenylthio” refers to —S-cycloalkenyl.

“Substituted cycloalkenylthio” refers to —S-(substituted cycloalkenyl).

“Guanidino” refers to the group —NHC(═NH)NH₂.

“Substituted guanidino” refers to —NR⁵³C(═NR⁵³)N(R⁵³)₂ where each R⁵³ isindependently selected from the group consisting of hydrogen, alkyl,substituted alkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclic, andsubstituted heterocyclic and two R⁵³ groups attached to a commonguanidino nitrogen atom are optionally joined together with the nitrogenbound thereto to form a heterocyclic or substituted heterocyclic group,provided that at least one R⁵³ is not hydrogen, and wherein saidsubstituents are as defined herein.

“Halo” or “halogen” refers to fluoro, chloro, bromo and iodo.

“Hydroxy” or “hydroxyl” refers to the group —OH.

“Heteroaryl” refers to an aromatic group of from 1 to 10 carbon atomsand 1 to 4 heteroatoms selected from the group consisting of oxygen,nitrogen and sulfur within the ring. Such heteroaryl groups can have asingle ring (e.g., pyridinyl or furyl) or multiple condensed rings(e.g., indolizinyl or benzothienyl) wherein the condensed rings may ormay not be aromatic and/or contain a heteroatom provided that the pointof attachment is through an atom of the aromatic heteroaryl group. Inone embodiment, the nitrogen and/or the sulfur ring atom(s) of theheteroaryl group are optionally oxidized to provide for the N-oxide(N→O), sulfinyl, or sulfonyl moieties. Certain non-limiting examplesinclude pyridinyl, pyrrolyl, indolyl, thiophenyl, oxazolyl, thizolyl,and furanyl.

“Substituted heteroaryl” refers to heteroaryl groups that aresubstituted with from 1 to 5, preferably 1 to 3, or more preferably 1 to2 substituents selected from the group consisting of the same group ofsubstituents defined for substituted aryl.

“Heteroaryloxy” refers to —O-heteroaryl.

“Substituted heteroaryloxy” refers to the group —O-(substitutedheteroaryl).

“Heteroarylthio” refers to the group —S-heteroaryl.

“Substituted heteroarylthio” refers to the group —S-(substitutedheteroaryl).

“Heterocycle” or “heterocyclic” or “heterocycloalkyl” or “heterocyclyl”refers to a saturated or partially saturated, but not aromatic, grouphaving from 1 to 10 ring carbon atoms and from 1 to 4 ring heteroatomsselected from the group consisting of nitrogen, sulfur, or oxygen.Heterocycle encompasses single ring or multiple condensed rings,including fused bridged and spiro ring systems. In fused ring systems,one or more the rings can be cycloalkyl, aryl, or heteroaryl providedthat the point of attachment is through a non-aromatic ring. In oneembodiment, the nitrogen and/or sulfur atom(s) of the heterocyclic groupare optionally oxidized to provide for the N-oxide, sulfinyl, orsulfonyl moieties.

“Substituted heterocyclic” or “substituted heterocycloalkyl” or“substituted heterocyclyl” refers to heterocyclyl groups that aresubstituted with from 1 to 5 or preferably 1 to 3 of the samesubstituents as defined for substituted cycloalkyl.

“Heterocyclyloxy” refers to the group —O-heterocycyl.

“Substituted heterocyclyloxy” refers to the group —O-(substitutedheterocycyl).

“Heterocyclylthio” refers to the group —S-heterocycyl.

“Substituted heterocyclylthio” refers to the group —S-(substitutedheterocycyl).

Examples of heterocycle and heteroaryls include, but are not limited to,azetidine, pyrrole, furan, thiophene, imidazole, pyrazole, pyridine,pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole,dihydroindole, indazole, purine, quinolizine, isoquinoline, quinoline,phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline,pteridine, carbazole, carboline, phenanthridine, acridine,phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine,phenothiazine, imidazolidine, imidazoline, piperidine, piperazine,indoline, phthalimide, 1,2,3,4-tetrahydroisoquinoline,4,5,6,7-tetrahydrobenzo[b]thiophene, thiazole, thiazolidine, thiophene,benzo[b]thiophene, morpholinyl, thiomorpholinyl (also referred to asthiamorpholinyl), 1,1-dioxothiomorpholinyl, piperidinyl, pyrrolidine,and tetrahydrofuranyl.

“Nitro” refers to the group —NO₂.

“Oxo” refers to the atom (═O).

Phenylene refers to a divalent aryl ring, where the ring contains 6carbon atoms.

Substituted phenylene refers to phenylenes which are substituted with 1to 4, preferably 1 to 3, or more preferably 1 to 2 substituents selectedfrom the group consisting of alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substitutedalkoxy, acyl, acylamino, acyloxy, amino, substituted amino,aminocarbonyl, aminothiocarbonyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, aminosulfonyl,aminosulfonyloxy, aminosulfonylamino, amidino, aryl, substituted aryl,aryloxy, substituted aryloxy, arylthio, substituted arylthio, carboxyl,carboxyl ester, (carboxyl ester)amino, (carboxyl ester)oxy, cyano,cycloalkyl, substituted cycloalkyl, cycloalkyloxy, substitutedcycloalkyloxy, cycloalkylthio, substituted cycloalkylthio, cycloalkenyl,substituted cycloalkenyl, cycloalkenyloxy, substituted cycloalkenyloxy,cycloalkenylthio, substituted cycloalkenylthio, guanidino, substitutedguanidino, halo, hydroxy, heteroaryl, substituted heteroaryl,heteroaryloxy, substituted heteroaryloxy, heteroarylthio, substitutedheteroarylthio, heterocyclic, substituted heterocyclic, heterocyclyloxy,substituted heterocyclyloxy, heterocyclylthio, substitutedheterocyclylthio, nitro, SO₃H, substituted sulfonyl, substitutedsulfonyloxy, thioacyl, thiol, alkylthio, and substituted alkylthio,wherein said substituents are as defined herein.

“Spirocycloalkyl” and “spiro ring systems” refers to divalent cyclicgroups from 3 to 10 carbon atoms having a cycloalkyl or heterocycloalkylring with a spiro union (the union formed by a single atom which is theonly common member of the rings) as exemplified by the followingstructure:

“Sulfonyl” refers to the divalent group —S(O)₂—.

“Substituted sulfonyl” refers to the group —SO₂-alkyl, —SO₂-substitutedalkyl, —SO₂-alkenyl, —SO₂-substituted alkenyl, —SO₂-cycloalkyl,—SO₂-substituted cylcoalkyl, —SO₂-cycloalkenyl, —SO₂-substitutedcylcoalkenyl, —SO₂-aryl, —SO₂-substituted aryl, —SO₂-heteroaryl,—SO₂-substituted heteroaryl, —SO₂-heterocyclic, —SO₂-substitutedheterocyclic, wherein alkyl, substituted alkyl, alkenyl, substitutedalkenyl, alkynyl, substituted alkynyl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, heterocyclic, and substitutedheterocyclic are as defined herein. Substituted sulfonyl includes groupssuch as methyl-SO₂—, phenyl-SO₂—, and 4-methylphenyl-SO₂—.

“Substituted sulfonyloxy” refers to the group —OSO₂-alkyl,—OSO₂-substituted alkyl, —OSO₂-alkenyl, —OSO₂-substituted alkenyl,—OSO₂-cycloalkyl, —OSO₂-substituted cylcoalkyl, —OSO₂-cycloalkenyl,—OSO₂-substituted cylcoalkenyl, —OSO₂-aryl, —OSO₂-substituted aryl,—OSO₂-heteroaryl, —OSO₂-substituted heteroaryl, —OSO₂-heterocyclic,—OSO₂-substituted heterocyclic, wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic are as defined herein.

“Thioacyl” refers to the groups H—C(S)—, alkyl-C(S)—, substitutedalkyl-C(S)—, alkenyl-C(S)—, substituted alkenyl-C(S)—, alkynyl-C(S)—,substituted alkynyl-C(S)—, cycloalkyl-C(S)—, substitutedcycloalkyl-C(S)—, cycloalkenyl-C(S)—, substituted cycloalkenyl-C(S)—,aryl-C(S)—, substituted aryl-C(S)—, heteroaryl-C(S)—, substitutedheteroaryl-C(S)—, heterocyclic-C(S)—, and substitutedheterocyclic-C(S)—, wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic are as defined herein.

“Thiol” refers to the group —SH.

“Thiocarbonyl” refers to the divalent group —C(S)— which is equivalentto —C(═S)—.

“Thioxo” refers to the atom (═S).

“Alkylthio” refers to the group —S-alkyl wherein alkyl is as definedherein.

“Substituted alkylthio” refers to the group —S-(substituted alkyl)wherein substituted alkyl is as defined herein.

A substituted ring can be substituted with one or more fused and/orspiro cycles. Such fused cycles include a fused cycloalkyl, a fusedheterocyclyl, a fused aryl, a fused heteroaryl ring, each of which ringscan be unsubstituted or substituted. Such spiro cycles include a fusedcycloalkyl and a fused heterocyclyl, each of which rings can beunsubstituted or substituted.

“Optionally substituted” refers to a group selected from that group anda substituted form of that group. Substituents are such as those definedhereinabove. In one embodiment, substituents are selected from C₁-C₁₀ orC₁-C₆ alkyl, substituted C₁-C₁₀ or C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, C₆-C₁₀ aryl, C₃-C₈ cycloalkyl, C₂-C₁₀ heterocyclyl, C₁-C₁₀heteroaryl, substituted C₂-C₆ alkenyl, substituted C₂-C₆ alkynyl,substituted C₆-C₁₀ aryl, substituted C₃-C₈ cycloalkyl, substitutedC₂-C₁₀ heterocyclyl, substituted C₁-C₁₀ heteroaryl, halo, nitro, cyano,—CO₂H or a C₁-C₆ alkyl ester thereof.

Unless indicated otherwise, the nomenclature of substituents that arenot explicitly defined herein are arrived at by naming the terminalportion of the functionality followed by the adjacent functionalitytoward the point of attachment. For example, the substituent“alkoxycarbonylalkyl” refers to the group (alkoxy)-C(O)-(alkyl)-.

It is understood that in all substituted groups defined above, polymersarrived at by defining substituents with further substituents tothemselves (e.g., substituted aryl having a substituted aryl group as asubstituent which is itself substituted with a substituted aryl group,etc.) are not intended for inclusion herein. In such cases, the maximumnumber of such substituents is three. That is to say that each of theabove definitions is constrained by a limitation that, for example,substituted aryl groups are limited to -substituted aryl-(substitutedaryl)-substituted aryl.

It is understood that the above definitions are not intended to includeimpermissible substitution patterns (e.g., methyl substituted with 5fluoro groups). Such impermissible substitution patterns are well knownto the skilled artisan.

“Tautomer” refer to alternate forms of a compound that differ in theposition of a proton, such as enol-keto and imine-enamine tautomers, orthe tautomeric forms of heteroaryl groups containing a ring atomattached to both a ring —NH— moiety and a ring ═N— moiety such aspyrazoles, imidazoles, benzimidazoles, triazoles, and tetrazoles.

“Uracil isostere” refers to an isostere of uracil and does not includeuracil or any halouracil. Such a moiety provides some or all of thehydrogen bond acceptor-donor-acceptor property of uracil and optionallyprovides other structural characteristics of uracil. A skilled artisanwill further appreciate the meaning of this term by reading the nonlimiting examples of such uracil isosteres provided herein.

As used herein, the term stereochemically pure denotes a compound whichhas 80% or greater by weight of the indicated stereoisomer and 20% orless by weight of other stereoisomers. In a further embodiment, thecompound of Formula (I) has 90% or greater by weight of the statedstereoisomer and 10% or less by weight of other stereoisomers. In a yetfurther embodiment, the compound of Formula (I) has 95% or greater byweight of the stated stereoisomer and 5% or less by weight of otherstereoisomers. In a still further embodiment, the compound of formula(I) has 97% or greater by weight of the stated stereoisomer and 3% orless by weight of other stereoisomers.

“Pharmaceutically acceptable salt” refers to salts of a compound, whichsalts are suitable for pharmaceutical use and are derived from a varietyof organic and inorganic counter ions well known in the art and include,when the compound contains an acidic functionality, by way of exampleonly, sodium, potassium, calcium, magnesium, ammonium, andtetraalkylammonium; and when the molecule contains a basicfunctionality, salts of organic or inorganic acids, such ashydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, andoxalate (see Stahl and Wermuth, eds., “Handbook of PharmaceuticallyAcceptable Salts,” (2002), Verlag Helvetica Chimica Acta, Zurich,Switzerland), for a discussion of pharmaceutical salts, their selection,preparation, and use.

Generally, pharmaceutically acceptable salts are those salts that retainsubstantially one or more of the desired pharmacological activities ofthe parent compound and which are suitable for in vivo administration.Pharmaceutically acceptable salts include acid addition salts formedwith inorganic acids or organic acids. Inorganic acids suitable forforming pharmaceutically acceptable acid addition salts include, by wayof example and not limitation, hydrohalide acids (e.g., hydrochloricacid, hydrobromic acid, hydroiodic acid, etc.), sulfuric acid, nitricacid, phosphoric acid, and the like.

Organic acids suitable for forming pharmaceutically acceptable acidaddition salts include, by way of example and not limitation, aceticacid, trifluoroacetic acid, propionic acid, hexanoic acid,cyclopentanepropionic acid, glycolic acid, oxalic acid, pyruvic acid,lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, tartaric acid, citric acid, palmitic acid, benzoic acid,3-(4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid,alkylsulfonic acids (e.g., methanesulfonic acid, ethanesulfonic acid,1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, etc.),arylsulfonic acids (e.g., benzenesulfonic acid, 4-chlorobenzenesulfonicacid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid,camphorsulfonic acid, etc.), glutamic acid, hydroxynaphthoic acid,salicylic acid, stearic acid, muconic acid, and the like.

Pharmaceutically acceptable salts also include salts formed when anacidic proton present in the parent compound is either replaced by ametal ion (e.g., an alkali metal ion, an alkaline earth metal ion, or analuminum ion) or by an ammonium ion (e.g., an ammonium ion derived froman organic base, such as, ethanolamine, diethanolamine, triethanolamine,morpholine, piperidine, dimethylamine, diethylamine, triethylamine, andammonia).

An “effective amount” is an amount sufficient to effect beneficial ordesired results. An effective amount can be administered in one or moreadministrations, applications or dosages. Such delivery is dependent ona number of variables including the time period for which the individualdosage unit is to be used, the bioavailability of the therapeutic agent,the route of administration, etc. It is understood, however, thatspecific dose levels of the therapeutic agents disclosed herein for anyparticular subject depends upon a variety of factors including theactivity of the specific compound employed, bioavailability of thecompound, the route of administration, the age of the animal and itsbody weight, general health, sex, the diet of the animal, the time ofadministration, the rate of excretion, the drug combination, and theseverity of the particular disorder being treated and form ofadministration. In general, one will desire to administer an amount ofthe compound that is effective to achieve a serum level commensuratewith the concentrations found to be effective in vivo. Theseconsiderations, as well as effective formulations and administrationprocedures are well known in the art and are described in standardtextbooks.

“Therapeutically effective amount” of a drug or an agent refers to anamount of the drug or the agent that is an amount sufficient to obtain apharmacological response such as inhibiting dUTPase; or alternatively,is an amount of the drug or agent that, when administered to a patientwith a specified disorder or disease, is sufficient to have the intendedeffect, e.g., treatment, alleviation, amelioration, palliation orelimination of one or more manifestations of the specified disorder ordisease in the patient. A therapeutic effect does not necessarily occurby administration of one dose, and may occur only after administrationof a series of doses. Thus, a therapeutically effective amount may beadministered in one or more administrations.

As used herein, “treating” or “treatment” of a disease in a patientrefers to (1) preventing the symptoms or disease from occurring in ananimal that is predisposed or does not yet display symptoms of thedisease; (2) inhibiting the disease or arresting its development; or (3)ameliorating or causing regression of the disease or the symptoms of thedisease. As understood in the art, “treatment” is an approach forobtaining beneficial or desired results, including clinical results. Forthe purposes of this technology, beneficial or desired results caninclude one or more, but are not limited to, alleviation or ameliorationof one or more symptoms, diminishment of extent of a condition(including a disease), stabilized (i.e., not worsening) state of acondition (including disease), delay or slowing of condition (includingdisease), progression, amelioration or palliation of the condition(including disease), states and remission (whether partial or total),whether detectable or undetectable.

“dUTPase” means any of the following, which are considered to besynonymous, “deoxyuridine triphosphate nucleotidohydrolase”,“deoxyuridine triphosphate pyrophosphatase”, “dUTP nucleotidohydrolase”,“dUTP pyrophosphatase”, and other equivalent nomenclature for thedUTPase enzyme. In one aspect, dUTPase intends DUT-N and DUT-M. In otheraspects, it is DUT-N only, or alternatively, DUT-M only. The amino acidand coding sequences for dUTPase are known in the art and disclosed inU.S. Pat. No. 5,962,246. Methods for expressing and screening forexpression level of the enzyme are disclosed in U.S. Pat. No. 5,962,246and Ladner et al. (US Patent Publ. No. 2011/0212467A1).

“DUT-N” means the nuclear form of dUTPase.

“DUT-M” means the mitochondrial or cytoplasmic form of dUTPase.

“dUTPase-directed therapy” intends therapeutics that target the dUTPasepathway, e.g., in the case of cancer, e.g. TS-directed therapies and thefluoropyrimidines (such as 5-FU), pemetrexed (Alimta®), capecitabine(Xeloda®), S-1 and antifolates (such as methotrexate) and chemicalequivalents thereof. Non-limiting examples include 5-flurouracil (5-FU),TS-directed therapies and 5-FU based adjuvant therapy. Combinationtherapies can include any intervention that alters nucleotide poolsand/or sensitizes the immune cells or viruses to the dUTPase inhibitor,as are well known to the skilled artisan. For rheumatoid arthritis, forexample, the combination can be with an dihydrofolate reductase (DHFR)inhibitor such as methotrexate.

5-fluorouracil (5-FU) belongs to the family of therapy drugs calledpyrimidine based anti-metabolites. It is a pyrimidine analog, which istransformed into different cytotoxic metabolites that are thenincorporated into DNA and RNA thereby inducing cell cycle arrest andapoptosis. Chemical equivalents are pyrimidine analogs which result indisruption of DNA replication. Chemical equivalents inhibit cell cycleprogression at S phase resulting in the disruption of cell cycle andconsequently apoptosis. Equivalents to 5-FU include prodrugs, analogsand derivative thereof such as 5′-deoxy-5-fluorouridine(doxifluoroidine), 1-tetrahydrofuranyl-5-fluorouracil (ftorafur),capecitabine (Xeloda®), S-1 (MBMS-247616, consisting of tegafur and twomodulators, a 5-chloro-2,4-dihydroxypyridine and potassium oxonate),ralititrexed (tomudex), nolatrexed (Thymitaq, AG337), LY231514 andZD9331, as described for example in Papamicheal (1999) The Oncologist4:478-487.

“5-FU based adjuvant therapy” refers to 5-FU alone or alternatively thecombination of 5-FU with other treatments, that include, but are notlimited to radiation, methyl-CCNU, leucovorin, oxaliplatin, irinotecin,mitomycin, cytarabine, levamisole. Specific treatment adjuvant regimensare known in the art as FOLFOX, FOLFOX4, FOLFIRI, MOF (semustine(methyl-CCNU), vincrisine (Oncovin®) and 5-FU). For a review of thesetherapies see Beaven and Goldberg (2006) Oncology 20(5):461-470. Anexample of such is an effective amount of 5-FU and Leucovorin. Otherchemotherapeutics can be added, e.g., oxaliplatin or irinotecan.

Capecitabine is a prodrug of (5-FU) that is converted to its active formby the tumor-specific enzyme PynPase following a pathway of threeenzymatic steps and two intermediary metabolites,5′-deoxy-5-fluorocytidine (5′-DFCR) and 5′-deoxy-5-fluorouridine(5′-DFUR). Capecitabine is marketed by Roche under the trade nameXeloda®.

Leucovorin (Folinic acid) is an adjuvant used in cancer therapy. It isused in synergistic combination with 5-FU to improve efficacy of thechemotherapeutic agent. Without being bound by theory, addition ofLeucovorin is believed to enhance efficacy of 5-FU by inhibitingthymidylate synthase. It has been used as an antidote to protect normalcells from high doses of the anticancer drug methotrexate and toincrease the antitumor effects of fluorouracil (5-FU) andtegafur-uracil. It is also known as citrovorum factor and Wellcovorin.This compound has the chemical designation of L-Glutamic acidN[4[[(2-amino-5-formyl1,4,5,6,7,8hexahydro4oxo6-pteridinyl)methyl]amino]b-enzoyl],calcium salt (1:1).

“Oxaliplatin” (Eloxatin) is a platinum-based chemotherapy drug in thesame family as cisplatin and carboplatin. It is typically administeredin combination with fluorouracil and leucovorin in a combination knownas FOLFOX for the treatment of colorectal cancer. Compared to cisplatin,the two amine groups are replaced by cyclohexyldiamine for improvedantitumor activity. The chlorine ligands are replaced by the oxalatobidentate derived from oxalic acid in order to improve water solubility.Equivalents to Oxaliplatin are known in the art and include, but are notlimited to cisplatin, carboplatin, aroplatin, lobaplatin, nedaplatin,and JM-216 (see McKeage et al. (1997) J. Clin. Oncol. 201:1232-1237 andin general, Chemotherapy for Gynecological Neoplasm, Curr. Therapy andNovel Approaches, in the Series Basic and Clinical Oncology, Angioli etal. Eds., 2004).

“FOLFOX” is an abbreviation for a type of combination therapy that isused to treat cancer. This therapy includes 5-FU, oxaliplatin andleucovorin. “FOLFIRI” is an abbreviation for a type of combinationtherapy that is used treat cancer and comprises, or alternativelyconsists essentially of, or yet further consists of 5-FU, leucovorin,and irinotecan. Information regarding these treatments are available onthe National Cancer Institute's web site, cancer.gov, last accessed onJan. 16, 2008.

Irinotecan (CPT-11) is sold under the trade name of Camptosar. It is asemi-synthetic analogue of the alkaloid camptothecin, which is activatedby hydrolysis to SN-38 and targets topoisomerase I. Chemical equivalentsare those that inhibit the interaction of topoisomerase I and DNA toform a catalytically active topoisomerase I-DNA complex. Chemicalequivalents inhibit cell cycle progression at G2-M phase resulting inthe disruption of cell proliferation.

The term “adjuvant” therapy refers to administration of a therapy orchemotherapeutic regimen to a patient after removal of a tumor bysurgery. Adjuvant therapy is typically given to minimize or prevent apossible cancer reoccurrence. Alternatively, “neoadjuvant” therapyrefers to administration of therapy or chemotherapeutic regimen beforesurgery, typically in an attempt to shrink the tumor prior to a surgicalprocedure to minimize the extent of tissue removed during the procedure.

The phrase “first line” or “second line” or “third line” etc., refers tothe order of treatment received by a patient. First line therapyregimens are treatments given first, whereas second or third linetherapy are given after the first line therapy or after the second linetherapy, respectively. The National Cancer Institute defines first linetherapy as “the first treatment for a disease or condition. In patientswith cancer, primary treatment can be surgery, chemotherapy, radiationtherapy, or a combination of these therapies. First line therapy is alsoreferred to those skilled in the art as primary therapy and primarytreatment.” See National Cancer Institute website at www.cancer.gov,last visited on May 1, 2008. Typically, a patient is given a subsequentchemotherapy regimen because the patient did not shown a positiveclinical or sub-clinical response to the first line therapy or the firstline therapy has stopped.

As used herein, the term “antifolate” intends a drug or biologic thatimpairs the function of folic acids, e.g., an antimetabolite agent thatinhibits the use of a metabolite, i.e. another chemical that is part ofnormal metabolism. In cancer treatment, antimetabolites interfere withDNA production, thus cell division and growth of the tumor. Non-limitingexamples of these agents are dihydrofolate reductase inhibitors, such asmethotrexate, Aminopterin, and Pemetrexed; thymidylate synthaseinhibitors, such as Raltitrexed or Pemetrexed; purine based, i.e. anadenosine deaminase inhibitor, such as Pentostatin, a thiopurine, suchas Thioguanine and Mercaptopurine, a halogenated/ribonucleotidereductase inhibitor, such as Cladribine, Clofarabine, Fludarabine, or aguanine/guanosine: thiopurine, such as Thioguanine; or Pyrimidine based,i.e. cytosine/cytidine: hypomethylating agent, such as Azacitidine andDecitabine, a DNA polymerase inhibitor, such as Cytarabine, aribonucleotide reductase inhibitor, such as Gemcitabine, or athymine/thymidine: thymidylate synthase inhibitor, such as aFluorouracil (5-FU).

In one aspect, the term “chemical equivalent” means the ability of thechemical to selectively interact with its target protein, DNA, RNA orfragment thereof as measured by the inactivation of the target protein,incorporation of the chemical into the DNA or RNA or other suitablemethods. Chemical equivalents include, but are not limited to, thoseagents with the same or similar biological activity and include, withoutlimitation a pharmaceutically acceptable salt or mixtures thereof thatinteract with and/or inactivate the same target protein, DNA, or RNA asthe reference chemical.

The terms “oligonucleotide” or “polynucleotide” or “portion,” or“segment” thereof refer to a stretch of polynucleotide residues which islong enough to use in PCR or various hybridization procedures toidentify or amplify identical or related parts of mRNA or DNA molecules.The polynucleotide compositions of this invention include RNA, cDNA,genomic DNA, synthetic forms, and mixed polymers, both sense andantisense strands, and may be chemically or biochemically modified ormay contain non-natural or derivatized nucleotide bases, as will bereadily appreciated by those skilled in the art. Such modificationsinclude, for example, labels, methylation, substitution of one or moreof the naturally occurring nucleotides with an analog, internucleotidemodifications such as uncharged linkages (e.g., methyl phosphonates,phosphotriesters, phosphoamidates, carbamates, etc.), charged linkages(e.g., phosphorothioates, phosphorodithioates, etc.), pendent moieties(e.g., polypeptides), intercalators (e.g., acridine, psoralen, etc.),chelators, alkylators, and modified linkages (e.g., alpha anomericnucleic acids, etc.). Also included are synthetic molecules that mimicpolynucleotides in their ability to bind to a designated sequence viahydrogen bonding and other chemical interactions. Such molecules areknown in the art and include, for example, those in which peptidelinkages substitute for phosphate linkages in the backbone of themolecule.

When a genetic marker, e.g., over expression of dUTPase, is used as abasis for selecting a patient for a treatment described herein, thegenetic marker is measured before and/or during treatment, and thevalues obtained are used by a clinician in assessing any of thefollowing: (a) probable or likely suitability of an individual toinitially receive treatment(s); (b) probable or likely unsuitability ofan individual to initially receive treatment(s); (c) responsiveness totreatment; (d) probable or likely suitability of an individual tocontinue to receive treatment(s); (e) probable or likely unsuitabilityof an individual to continue to receive treatment(s); (f) adjustingdosage; (g) predicting likelihood of clinical benefits; or (h) toxicity.As would be well understood by one in the art, measurement of thegenetic marker in a clinical setting is a clear indication that thisparameter was used as a basis for initiating, continuing, adjustingand/or ceasing administration of the treatments described herein.

“Cancer” is a known medically as a malignant neoplasm, is a broad groupof diseases involving unregulated cell growth. In cancer, cells divideand grow uncontrollably, forming malignant tumors, and invade nearbyparts of the body. Non-limiting examples include colon cancer,colorectal cancer, gastric cancer, esophogeal cancer, head and neckcancer, breast cancer, lung cancer, stomach cancer, liver cancer, gallbladder cancer, or pancreatic cancer or leukemia.

Compounds

In one aspect, provided herein is a compound of formula (I):

or a tautomer thereof, or a pharmaceutically acceptable salt of eachthereof, or a pharmaceutically acceptable solvate of each of theforegoing, wherein

A is as defined above;

L¹ is as defined above;

L² is as defined above;

L³ is as defined above;

B is as defined above.

In some embodiments, B is

each R¹-R³ independently is H, F, Cl, C₁-C₃ alkyl, or OR²⁰;

R²⁰ is CH₂-R²¹; methyl optionally substituted with 2 or 3 fluorineatoms; C₃-C₆ cycloalkyl; or C₁-C₆ alkyl;

R²¹ is C₁-C₁₀ alkyl, preferably branched C₃-C₁₀ alkyl, more preferablyisopropyl or t-butyl, optionally substituted with one or more hydroxy orfluoro; C₃-C₆ cycloalkyl, preferably cyclopropyl, cyclobutyl,cyclopentyl; or

wherein each R²²-R²⁴ independently is an optionally substituted C₁-C₃alkyl or hydroxy.

In one embodiment, Y¹ is H. In another embodiment, Y¹ is C₁-C₃ alkyl.

In one embodiment, L¹ is an optionally substituted C₃-C₁₀ alkylene,further wherein at least two geminal hydrogens are optionallysubstituted with cyclopropano or cyclobutano. In another embodiment, L¹is an optionally substituted C₃-C₁₀ alkenylene. In yet anotherembodiment, L¹ is an optionally substituted C₃-C₁₀ heteroalkylene. In afurther embodiment, L¹ is an optionally substituted C₃-C₁₀heteroalkenylene.

In one embodiment, L¹ is -L¹¹-L¹²-L¹³-, wherein L¹¹ is attached to A. Inone embodiment, L¹¹ is O. In another embodiment, L¹¹ is S. In yetanother embodiment, L¹¹ is NR. In one embodiment, and R is H. In anotherembodiment, R is C₁-C₃ alkyl.

In one embodiment, L¹¹ is C₁-C₂ alkylene. In one embodiment, L¹¹ is C₂alkenylene. In another embodiment, L¹¹ is C₂ heteroalkylene. In yetanother embodiment, L¹¹ is C₃ heteroalkenylene.

In one embodiment, L¹² is arylene. In another embodiment, L¹² isheteroarylene.

In one embodiment, L¹³ is a bond. In another embodiment, L¹³ is anoptionally substituted C₁-C₅ alkylene.

In some embodiments, L² is —C(O)—. In some embodiments, L² is —S(O)₂—.

In some embodiments, L³ is a bond. In some embodiments, L³ is—C(R³⁰⁰)₂—.

In some embodiments, each R³⁰⁰ independently is a hydrogen, hydroxyl, oran optionally substituted phenyl. In some embodiments, each R³⁰⁰independently is a hydrogen. In some embodiments, each R³⁰⁰independently is a hydroxyl. In some embodiments, each R³⁰⁰independently is an optionally substituted phenyl.

In some embodiments, one R³⁰⁰ is hydroxyl and the second R³⁰⁰ is anoptionally substituted phenyl. In some embodiments, one R³⁰⁰ is hydrogenand the second R³⁰⁰ is an optionally substituted phenyl. In someembodiments, one R³⁰⁰ is a phenyl, wherein the phenyl is notsubstituted.

In one embodiment, each R¹-R³ independently is H. In one embodiment,each R¹-R³ independently is F. In one embodiment, each 10-R³independently is Cl. In one embodiment, each R¹-R³ independently isC₁-C₃ alkyl. In one embodiment, each R¹-R³ independently is OR²⁰.

In one embodiment, R²⁰ is CH₂-R²¹. In one embodiment, R²⁰ is methyloptionally substituted with 2 or 3 fluorine atoms. In one embodiment,R²⁰ is C₃-C₆ cycloalkyl. In one embodiment, R²⁰ is C₁-C₆ alkyl.

In one embodiment, R²¹ is C₁-C₁₀ alkyl. In one embodiment, R²¹ is abranched C₃-C₁₀ alkyl optionally substituted with one or more hydroxy orfluoro. In another embodiment, R²¹ is isopropyl or t-butyl optionallysubstituted with one or more hydroxy or fluoro. In another embodiment,R²¹ is a C₃-C₆ cycloalkyl. In yet another embodiment, R²¹ is acyclopropyl, cyclobutyl, or cyclopentyl.

In one embodiment, R²¹ is

wherein each R²²-R²⁴ independently is an optionally substituted C₁-C₃alkyl or hydroxy.

In one embodiment, each R²²-R²⁴ independently is an optionallysubstituted C₁-C₃ alkyl. In another embodiment, each R²²-R²⁴independently is a hydroxy.

In one embodiment, R²¹ is

In one embodiment, R²¹ is

In one embodiment, R²¹ is

In one embodiment, R²¹ is

In one embodiment, R²¹ is

In one embodiment, R²¹ is

In one embodiment, R²¹ is

In one embodiment, R²¹ is

In one embodiment, A is selected from the group consisting of:

In one embodiment, A is

In one embodiment, A is

In one embodiment, A is

In one embodiment, L¹ is

—(CH₂)_(q)—, wherein one or more hydrogens are optionally substitutedwith C₁-C₃ alkyl and/or at least two or more geminal hydrogens areoptionally substituted with cyclopropano or cyclobutano; and wherein qis 4, 5, 6, 7, or 8.

In another embodiment, L¹ is

wherein one or more hydrogens are optionally substituted with C₁-C₃alkyl and/or at least two or more geminal hydrogens are optionallysubstituted with cyclopropano or cyclobutano; and wherein p is 0, 1, 2,3, 4, or 5 and z is 0, 1, 2, 3, 4, or 5.

In yet another embodiment, L¹ is

—(CH₂)_(m)—X—(CH₂)_(n)—, wherein one or more hydrogens are optionallysubstituted with C₁-C₃ alkyl and/or at least two or more geminalhydrogens are optionally substituted with cyclopropano or cyclobutano;and wherein m is 0, 1, 2, or 3 and n is 3, 4, 5, 6, or 7.

In a further embodiment, L¹ is

wherein one or more hydrogens are optionally substituted with C₁-C₃alkyl and/or at least two or more geminal hydrogens are optionallysubstituted with cyclopropano or cyclobutano; and wherein o is 0, 1, 2,or 3; r is 1, 2 or 3; and s is 0, 1, 2, 3, or 4; and

wherein X is NR⁴⁰, O, or S, wherein R⁴⁰ is H or C₁-C₃ alkyl.

In one embodiment, L¹ is selected from the group consisting of:

wherein the left side of the moieties are attached to A.

In another embodiment, -L¹¹-L¹²-L¹³- is

wherein the left side of the moieties are attached to A.

In one embodiment, R¹ is H.

In one embodiment, R² is H or —OR²⁰.

In one embodiment, R³ is F or H.

In one embodiment, B is

In one embodiment, B is selected from the group consisting of:

In one embodiment, B is

In one aspect, provided herein is a compound selected from Table 1Abelow. In another aspect, provided herein is a compound selected fromTable 3 below. In another aspect, provided herein is a compound selectedfrom Table 1C below. In another aspect, provided herein is a compoundselected from Table 1A or Table 1B below. In another aspect, providedherein is a compound selected from Table 1A, Table 1B, or Table 1Cbelow.

TABLE lA 1222

1223

1224

1225

1226

1227

1229

1230

1231

1232

1233

1234

1235

1236

1237

1238

1239

TABLE 1B  81

1138

1149

1151

1152

1153

1155

1156

1165

1177

1178

1180

1186

 12

TABLE 1C 1914

1915

1916

1917

1918

Synthesis

These and other compounds provided herein are synthesized following artrecognized methods with the appropriate substitution of commerciallyavailable reagents as needed. For example, and without limitation,methods for synthesizing certain other compounds are described in US2011/0082163; US 2012/0225838; WO 2014/107622; PCT/US2015/010059; US2016/0039788; US 2016/0326149; PCT/IB2016/054091, PCT/IB2016/054092,PCT/IB2016/054067, PCT/IB2016/054069, Miyahara et al., J. Med. Chem.(2012) 55, 2970-2980; Miyakoshi et al., J. Med. Chem. (2012) 55,2960-2969; Miyahara et al., J. Med. Chem. (2012) 55 (11), pp 5483-5496;and Miyakoshi et al., J. Med. Chem. (2012) 55 (14), pp 6427-6437 (eachsupra), which methods can be adapted by the skilled artisan upon readingthis disclosure and/or based on synthetic methods well known in the art,to prepare the compounds provided herein. Protection deprotectionmethods and protecting groups useful for such purposes are well known inthe art, for example in Greene's Protective Groups in Organic Synthesis,4^(th) Edition, Wiley, 2006, or a later edition of the book.

The compounds and the intermediates are separated from the reactionmixture, when desired, following art known methods such ascrystallization, chromatography, distillation, and the like. Thecompounds and the intermediates are characterized by art known methodssuch as thin layer chromatography, nuclear magnetic resonancespectroscopy, high performance liquid chromatography, and the like. Asdescribed in detail herein, a racemic or diastereomeric mixture of thecompound can be separated or enriched to the enantiomers anddiastereomers and tested and used diagnostically or therapeutically asdescribed herein.

Methods of testing and using the compounds provided herein are performedfollowing art recognized in vitro (cell free), ex vivo or in vivomethods. For example, and without limitation, certain methods fortesting and using other compounds are described in US 2011/0082163; US2012/0225838; US 2016/0039788; US 2016/0326149; PCT/IB2016/054091,PCT/IB2016/054092, PCT/IB2016/054067, PCT/IB2016/054069, Miyahara etal., J. Med. Chem. (2012) 55, 2970-2980; Miyakoshi et al., J. Med. Chem.(2012) 55, 2960-2969; Miyahara et al., J. Med. Chem. (2012) 55 (11), pp5483-5496; Miyakoshi et al., J. Med. Chem. (2012) 55 (14), pp 6427-6437(each of which in incorporated by reference), which methods can beadapted by the skilled artisan upon reading this disclosure and/or basedon methods well known in the art, to test and use the compounds providedherein.

Scheme A provides illustrative and non-limiting, generic routes for thesynthesis of the compounds provided herein. As is apparent, thevariables shown in Scheme A applies to Scheme A.

Pharmaceutical Compositions

In another aspect, provided herein is a composition comprising acompound provided herein, and at least one pharmaceutically acceptableexcipient.

Compositions, including pharmaceutical compositions comprising thecompounds described herein can be manufactured by means of conventionalmixing, dissolving, granulating, dragee-making levigating, emulsifying,encapsulating, entrapping, or lyophilization processes. The compositionscan be formulated in conventional manner using one or morephysiologically acceptable carriers, diluents, excipients, orauxiliaries which facilitate processing of the compounds provided hereininto preparations which can be used pharmaceutically.

The compounds of the technology can be administered by parenteral (e.g.,intramuscular, intraperitoneal, intravenous, ICV, intracisternalinjection or infusion, subcutaneous injection, or implant), oral, byinhalation spray nasal, vaginal, rectal, sublingual, urethral (e.g.,urethral suppository) or topical routes of administration (e.g., gel,ointment, cream, aerosol, etc.) and can be formulated, alone ortogether, in suitable dosage unit formulations containing conventionalnon-toxic pharmaceutically acceptable carriers, adjuvants, excipients,and vehicles appropriate for each route of administration.

In one embodiment, this technology relates to a composition comprising acompound as described herein and a carrier.

In another embodiment, this technology relates to a pharmaceuticalcomposition comprising a compound as described herein and apharmaceutically acceptable carrier.

In another embodiment, this technology relates to a pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundas described herein and a pharmaceutically acceptable carrier.

The pharmaceutical compositions for the administration of the compoundscan be conveniently presented in dosage unit form and can be prepared byany of the methods well known in the art of pharmacy. The pharmaceuticalcompositions can be, for example, prepared by uniformly and intimatelybringing the compounds provided herein into association with a liquidcarrier, a finely divided solid carrier or both, and then, if necessary,shaping the product into the desired formulation. In the pharmaceuticalcomposition the compound provided herein is included in an amountsufficient to produce the desired therapeutic effect. For example,pharmaceutical compositions of the technology may take a form suitablefor virtually any mode of administration, including, for example,topical, ocular, oral, buccal, systemic, nasal, injection, infusion,transdermal, rectal, and vaginal, or a form suitable for administrationby inhalation or insufflation.

For topical administration, the compounds can be formulated assolutions, gels, ointments, creams, suspensions, etc., as is well-knownin the art.

Systemic formulations include those designed for administration byinjection (e.g., subcutaneous, intravenous, infusion, intramuscular,intrathecal, or intraperitoneal injection) as well as those designed fortransdermal, transmucosal, oral, or pulmonary administration.

Useful injectable preparations include sterile suspensions, solutions,or emulsions of the compounds provided herein in aqueous or oilyvehicles. The compositions may also contain formulating agents, such assuspending, stabilizing, and/or dispersing agents. The formulations forinjection can be presented in unit dosage form, e.g., in ampules or inmultidose containers, and may contain added preservatives.

Alternatively, the injectable formulation can be provided in powder formfor reconstitution with a suitable vehicle, including but not limited tosterile pyrogen free water, buffer, and dextrose solution, before use.To this end, the compounds provided herein can be dried by any art-knowntechnique, such as lyophilization, and reconstituted prior to use.

For transmucosal administration, penetrants appropriate to the barrierto be permeated are used in the formulation. Such penetrants are knownin the art.

For oral administration, the pharmaceutical compositions may take theform of, for example, lozenges, tablets, or capsules prepared byconventional means with pharmaceutically acceptable excipients such asbinding agents (e.g., pregelatinised maize starch, polyvinylpyrrolidone,or hydroxypropyl methylcellulose); fillers (e.g., lactose,microcrystalline cellulose, or calcium hydrogen phosphate); lubricants(e.g., magnesium stearate, talc, or silica); disintegrants (e.g., potatostarch or sodium starch glycolate); or wetting agents (e.g., sodiumlauryl sulfate). The tablets can be coated by methods well known in theart with, for example, sugars, films, or enteric coatings.

Compositions intended for oral use can be prepared according to anymethod known to the art for the manufacture of pharmaceuticalcompositions, and such compositions may contain one or more agentsselected from the group consisting of sweetening agents, flavoringagents, coloring agents, and preserving agents in order to providepharmaceutically elegant and palatable preparations. Tablets contain thecompounds provided herein in admixture with non-toxic pharmaceuticallyacceptable excipients which are suitable for the manufacture of tablets.These excipients can be for example, inert diluents, such as calciumcarbonate, sodium carbonate, lactose, calcium phosphate or sodiumphosphate; granulating and disintegrating agents (e.g., corn starch oralginic acid); binding agents (e.g. starch, gelatin, or acacia); andlubricating agents (e.g., magnesium stearate, stearic acid, or talc).The tablets can be left uncoated or they can be coated by knowntechniques to delay disintegration and absorption in thegastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonostearate or glyceryl distearate can be employed. They may also becoated by the techniques well known to the skilled artisan. Thepharmaceutical compositions of the technology may also be in the form ofoil-in-water emulsions.

Liquid preparations for oral administration may take the form of, forexample, elixirs, solutions, syrups, or suspensions, or they can bepresented as a dry product for constitution with water or other suitablevehicle before use. Such liquid preparations can be prepared byconventional means with pharmaceutically acceptable additives such assuspending agents (e.g., sorbitol syrup, cellulose derivatives, orhydrogenated edible fats); emulsifying agents (e.g., lecithin, oracacia); non-aqueous vehicles (e.g., almond oil, oily esters, ethylalcohol, Cremophore™, or fractionated vegetable oils); and preservatives(e.g., methyl or propyl-p-hydroxybenzoates or sorbic acid). Thepreparations may also contain buffer salts, preservatives, flavoring,coloring, and sweetening agents as appropriate.

Use of Compounds for Preparing Medicaments

The compounds and compositions of the present invention are also usefulin the preparation of medicaments to treat a variety of pathologies asdescribed herein. The methods and techniques for preparing medicamentsof a composition are known in the art. For the purpose of illustrationonly, pharmaceutical formulations and routes of delivery are detailedherein.

Thus, one of skill in the art would readily appreciate that any one ormore of the compositions described above, including the many specificembodiments, can be used by applying standard pharmaceuticalmanufacturing procedures to prepare medicaments to treat the manydisorders described herein. Such medicaments can be delivered to thesubject by using delivery methods known in the pharmaceutical arts.

Methods of Treatment and Therapies

The compositions and compounds as disclosed herein are useful in methodsof inhibiting dUTPase or enhancing the efficacy of a dUTPase-directedtherapy, or yet further, reversing resistance to dUTPase therapies. Themethods comprise, or alternatively consist essentially of, or yetfurther consist of, contacting the dUTPase with a therapeuticallyeffective amount of the compound or composition as disclosed herein. Inone embodiment, the methods further comprise, or alternatively consistessentially of, or yet further consist of, contacting the dUTPase withan effective amount of a dUTPase-directed therapy. In one aspect, thecontacting of the dUTPase-directed therapy is prior to, concurrent orsubsequent to contacting with the compound or composition of thisdisclosure.

One of skill in the art can also determine if the compound orcombination inhibits dUTPase in vitro by contacting the compound orcombination with purified or recombinant dUTPase in a cell free system.The purified or recombinant dUTPase and can be from any species, e.g.,simian, canine, bovine, ovine, rat, mouse or human. In one aspect, thedUTPase is DUT-N or DUT-M. Isolation, characterization and expression ofdUTPase isoforms are disclosed in U.S. Pat. No. 5,962,246 and known inthe art.

The contacting can be performed cell-free in vitro or ex vivo with acell or in a cell culture. When performed in vitro or ex vivo, thecompounds, compositions or agents can be directly added to the enzymesolution or added to the cell culture medium. When practiced in vitro orex vivo, the method can be used to screen for novel combinationtherapies, formulations or treatment regimens, prior to administrationto administration to an animal or a human patient. Methods to quantifyinhibition are known in the art, see, U.S. Patent Publ. Nos.2010/0075924 and 2011/0212467 and U.S. Pat. No. 7,601,702. For example,a fixed dose of a dUTPase directed therapy (e.g., 5-FU or Pemetrexed)can be added to the system and varying amounts of the compound can besubsequently added to system. Alternatively, a fixed dose of a compoundof this invention can be added to the system and varying amounts of thedUTPase directed therapy (e.g., 5-FU or Pemetrexed) compound can besubsequently added to system.

In one aspect, the contacting is ex vivo and the cell or tissue to becontacted over expresses dUTPase. These cells can be isolated from apatient prior to administration to the patient or can be purchased froma depository such as the American Type Culture Collection (ATCC).Non-limiting examples of animal (e.g., canine, an equine, a bovine, afeline, an ovine, a mouse, a rat or a simian) and human cells that areknown to over express dUTPase include, without limitation cancer cells,e.g. colon cancer, colorectal cancer, gastric cancer, head and neckcancer, breast cancer, stomach cancer or lung cancer. The cancer can bemetastatic or non-metastatic. Methods to quantify inhibition are knownin the art, see, U.S. Patent Publ. Nos. 2010/0075924 and 2011/0212467and U.S. Pat. No. 7,601,702 and Wilson et al. (2012) Mol. Cancer Ther.11:616-628.

When practiced in vivo in a patient such as an animal or human, thecompounds, compositions or agents are administered in an effectiveamount by a suitable route of administration, as determined by atreating physician taking into account the patient, disease and otherfactors. When practiced in a non-human animal, e.g., an appropriatemouse model, the method can be used to screen for novel combinationtherapies, formulations or treatment regimens, prior to administrationto a human patient.

This disclosure also provides methods of treating a disease whosetreatment is impeded by the expression of dUTPase, comprising, oralternatively consisting essentially of, or yet further consisting of,administering to a patient in need of such treatment a therapeuticallyeffective amount of the compound or composition of this disclosure,thereby treating the disease. In one aspect, the method furthercomprises isolating a cell or tissue sample from the patient andscreening for the expression level of dUTPase, wherein over expressionof dUTPase in the sample as compared to a control sample serves as abasis for selecting the patient as suitable for the method andtherapies. Methods to quantify dUTPase are known in the art. Effectiveamounts will vary with the patient, the disease and the general healthof the patient and are determined by the treating physician. Methods toquantify inhibition are known in the art, see, U.S. Patent Publ. Nos.2010/0075924 and 2011/0212467 and U.S. Pat. No. 7,601,702 and Wilson etal. (2012) Mol. Cancer Ther. 11:616-628. If the patient sample showsover expression of dUTPase, the therapy is administered to the patient.If the patient sample does not show over expression, an alternatetherapy is chosen. The screen can be repeated throughout therapy as ameans to monitor the therapy and/or dosage regimen.

To practice this method, the sample is a patient sample containing thetumor tissue, normal tissue adjacent to said tumor, normal tissue distalto said tumor or peripheral blood lymphocytes. In a further aspect, thepatient or patient population to be treated also is treatment naïve.

In one aspect, the method also requires isolating a sample containingthe genetic material to be tested; however, it is conceivable that oneof skill in the art will be able to analyze and identify genetic markersin situ at some point in the future. Accordingly, in one aspect, theinventions of this application are not to be limited to requiringisolation of the genetic material prior to analysis.

These methods also are not limited by the technique that is used toidentify the expression level or in aspects where expression has beenlinked to a polymorphism, the polymorphism of interest. Suitable methodsinclude but are not limited to the use of hybridization probes,antibodies, primers for PCR analysis, and gene chips, slides andsoftware for high throughput analysis. Additional genetic markers can beassayed and used as negative controls.

In one aspect, the subject or patient is an animal or a human patient.Non-limiting examples of animals include a feline, a canine, a bovine,an equine, an ovine, a mouse, a rat or a simian.

Diseases in which treatment is impeded by the expression of dUTPaseinclude, without limitation, cancer, viral infection, bacterialinfection or an autoimmune disorder. For example, in inflammatory boweldisease or other autoimmune disorders, a dUTPase inhibitor can be usedin combination with an antifolate or fluoropyrimidine or otherthymidylate synthase and dihydrofolate reductase inhibitors; parasitic,viral or bacterial infections can be treated similarly employing acombination therapy including a dUTPase inhibitor. Non-limiting examplesof cancer include, colon cancer, colorectal cancer, gastric cancer, headand neck cancer, breast cancer, ovarian cancer, stomach cancer, lungcancer or a leukemia. The cancer can be metastatic or non-metastatic.

In another aspect, the compounds or compositions provided herein areuseful in methods of inhibiting the growth of a cancer cell. The methodscomprise, or alternatively consist essentially of, or yet furtherconsist of, contacting the cell with a therapeutically effective amountof the compounds or compositions disclosed herein and a therapeuticallyeffective amount of a dUTPase directed therapeutic, thereby inhibitingthe growth of the cancer cell.

In one embodiment, the cancer cell is selected from a colon cancer cell,a colorectal cancer cell, a gastric cancer cell, a head and neck cancercell, a breast cancer cell, a lung cancer cell or a blood cell.

In one aspect, the compound or composition is administered as one ormore of: a first line therapy or alternatively, a second line therapy, athird line therapy, or a fourth or subsequent line therapy toadministration of a dUPTase-directed therapy. Non-limiting examples ofdUTPase-directed therapies include an antimetabolite or afluoropyrmidine therapy or a 5-FU based adjuvant therapy or anequivalent or each thereof, such as 5-FU, tegafur, gimeracil, oteracilpotassium, capcitabine, 5-fluoro-2′-deoxyuridine, methotrexate, orpemetrexed or an equivalent of each thereof.

Certain compounds provided herein demonstrated substantial, such as, 1%to more than 100%, such as 100-140%, 100-200%, or 120-200%, dUTPaseinhibitory effect, an ability to inhibit dUTPase under conditionsdescribed herein below, and/or known to the skilled artisan, compared,for example, to a positive control:

In some embodiments, certain compounds provided herein demonstrate100-140%, dUTPase inhibitory effect, an ability to inhibit dUTPase underconditions described herein below, and/or known to the skilled artisan,compared, for example, to the positive control. In some embodiments,certain compounds provided herein demonstrate 120-200%, dUTPaseinhibitory effect, an ability to inhibit dUTPase under conditionsdescribed herein below, and/or known to the skilled artisan, compared,for example, to the positive control. In some embodiments, certaincompounds provided herein demonstrate 100-200%, dUTPase inhibitoryeffect, an ability to inhibit dUTPase under conditions described hereinbelow, and/or known to the skilled artisan, compared, for example, tothe positive control.Kits

The compounds and compositions, as described herein, can be provided inkits. The kits can further contain additional dUTPase inhibitors andoptionally, instructions for use. In a further aspect, the kit containsreagents and instructions to perform the screen to identify patientsmore likely to respond to the therapy as described above.

Screening Assays

This disclosure also provides screening assays to identify potentialtherapeutic agents of known and new compounds and combinations. Forexample, one of skill in the art can also determine if the compound orcombination inhibits dUTPase in vitro by contacting the compound orcombination with purified or recombinant dUTPase in a cell free system.The purified or recombinant dUTPase and can be from any species, e.g.,simian, canine, bovine, ovine, rat, mouse or human. In one aspect, thedUTPase is DUT-N or DUT-M. Isolation, characterization and expression ofdUTPase isoforms are disclosed in U.S. Pat. No. 5,962,246 and known inthe art.

The contacting can be performed cell-free in vitro or ex vivo with acell or in a cell culture. When performed in vitro or ex vivo, thecompounds, compositions or agents can be directly added to the enzymesolution or added to the cell culture medium. When practiced in vitro orex vivo, the method can be used to screen for novel combinationtherapies, formulations or treatment regimens, prior to administrationto administration to an animal or a human patient. Methods to quantifyinhibition are known in the art, see, U.S. Patent Publ. Nos.2010/0075924 and 2011/0212467 and U.S. Pat. No. 7,601,702. For example,a fixed dose of a dUTPase directed therapy (e.g., 5-FU or Pemetrexed)can be added to the system and varying amounts of the compound can besubsequently added to system. Alternatively, a fixed dose of a compoundof this invention can be added to the system and varying amounts of thedUTPase directed therapy (e.g., 5-FU or Pemetrexed) compound can besubsequently added to system.

In another aspect, the assay requires contacting a first samplecomprising suitable cells or tissue (“control sample”) with an effectiveamount of a composition of this invention and optionally a dUTPaseinhibitor, and contacting a second sample of the suitable cells ortissue (“test sample”) with the agent to be assayed and optionally adUTPase inhibitor. In one aspect, the cell or tissue over expressdUTPase. The inhibition of growth of the first and second cell samplesare determined. If the inhibition of growth of the second sample issubstantially the same or greater than the first sample, then the agentis a potential drug for therapy. In one aspect, substantially the sameor greater inhibition of growth of the cells is a difference of lessthan about 1%, or alternatively less than about 5% or alternatively lessthan about 10%, or alternatively greater than about 10%, oralternatively greater than about 20%, or alternatively greater thanabout 50%, or alternatively greater than about 90%. The contacting canbe in vitro or in vivo. Means for determining the inhibition of growthof the cells are well known in the art.

In a further aspect, the test agent is contacted with a third sample ofcells or tissue comprising normal counterpart cells or tissue to thecontrol (or alternatively cells that do not over express dUTPase) andtest samples and selecting agents that treat the second sample of cellsor tissue but does not adversely affect the third sample. For thepurpose of the assays described herein, a suitable cell or tissue isdescribed herein such as cancer or other diseases as described herein.Examples of such include, but are not limited to cancer cell or tissueobtained by biopsy, blood, breast cells, colon cells.

Efficacy of the test composition is determined using methods known inthe art which include, but are not limited to cell viability assays orapoptosis evaluation.

In yet a further aspect, the assay requires at least two cell types, thefirst being a suitable control cell.

The assays also are useful to predict whether a subject will be suitablytreated by this invention by delivering a composition to a samplecontaining the cell to be treated and assaying for treatment which willvary with the pathology or for screening for new drugs and combinations.In one aspect, the cell or tissue is obtained from the subject orpatient by biopsy. Applicants provide kits for determining whether apathological cell or a patient will be suitably treated by this therapyby providing at least one composition of this invention and instructionsfor use.

The test cells can be grown in small multi-well plates and is used todetect the biological activity of test compounds. For the purposes ofthis invention, the successful candidate drug will block the growth orkill the pathogen but leave the control cell type unharmed.

The following examples are included to demonstrate some embodiments ofthe disclosure. However, those of skill in the art should, in light ofthe present disclosure, appreciate that many changes can be made in thespecific embodiments which are disclosed and still obtain a like orsimilar result without departing from the spirit and scope of theinvention.

The following examples illustrate making certain portions of thecompounds provided herein and provide methods for making othercompounds, which methods can be adapted by the skilled artisan to makethe compounds of the present invention.

EXAMPLES Synthetic Examples Exemplary Procedure for the Preparation ofN-allylcyanamide (I)

To a stirred solution of prop-2-en-1-amine (3.0 g, 0.52 mmol) in Et₂O(50 mL), CNBr (3.33 g, 0.31 mmol) in Et₂O (50 mL) was added dropwise at0° C. and stirred at room temperature for 1 h. The progress of thereaction was monitored by TLC. After completion of the reaction, thereaction mixture was filtered and filtrate was washed with water anddried to afford I.

Yield: 1.7 g, 39.4%; ¹H NMR (400 MHz, Chloroform-d) δ 5.91-5.87 (m, 1H),5.40-5.26 (m, 2H), 3.72-3.68 (m, 2H), 3.53-3.43 (m, 1H).

Exemplary Procedure for the Preparation of ethylN-allyl-N-cyanoglycinate (II)

To a stirred solution of I (1.7 g, 20.7 mmol) in dry THF (8 mL), NaH(0.54 mL, 22.7 mmol) was added at 0° C. and stirred at room temperature.After 1 h, 3 (3.34 g, 20.7 mmol) in THF (8 mL) was added drop wise andstirred at room temperature for 1 h. The progress of the reaction wasmonitored by TLC. After completion of the reaction, the reaction mixturewas quenched with water then extracted with DCM. Combined DCM layer wasdried, concentrated under reduced pressure to afford II.

Yield: 2.7 g, 77.5%; NMR: ¹H NMR (400 MHz, Chloroform-d) δ 5.88-5.84 (m,1H), 5.40-5.28 (m, 2H), 4.25 (d, J=7.0 Hz, 2H), 3.80-3.73 (m, 4H),0.88-0.85 (m, 3H).

Exemplary Procedure for the Preparation of1-allylimidazolidine-2,4-dione (III)

To a stirred solution of II (2.70 g, 16.0 mmol) in Et₂O (27 mL), 50%H₂SO₄ (13.5 mL) was added drop wise at 0° C., the reaction mixture wasstirred at same temperature for 30 min and allowed to room temperaturefor 7 h. The progress of the reaction was monitored by TLC. Aftercompletion of the reaction, the reaction mixture was with ice cold waterand precipitated solid was filtrated and washed with ether to affordIII.

Yield: 0.1 g, 7.14%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.78 (s, 1H),5.90-5.69 (m, 1H), 5.25-5.05 (m, 2H), 3.91-3.79 (m, 4H).

Exemplary Procedure for the Preparation of but-3-ene-1-sulfonic acid(IV)

To a stirred solution of 4-bromobut-1-ene (4.00 g, 33.05 mmol) in water(30 mL), Na₂SO₃ (8.33 g, 66.11 mmol) was added and heated at 100° C. for16 h. The progress of the reaction was monitored by TLC. Aftercompletion of the reaction, the reaction mixture was quenched with waterand washed with ether. Aqueous layer was concentrated to dryness toafford the title compound IV.

Yield: 7 g, crude; ¹H NMR (400 MHz, D₂O) δ 5.97-5.95 (m, 1H), 5.29-5.06(m, 2H), 3.17-2.89 (m, 2H), 2.57-2.46 (m, 2H).

Exemplary Procedure for the Preparation of but-3-ene-1-sulfonyl chloride(V)

To a stirred solution of IV (7 g, 48.61 mmol) in (COCl)₂ (70 mL), DMF(1.5 mL) was added at 0° C. and stirred at room temperature for 3 h. Theprogress of the reaction was monitored by TLC. After completion of thereaction, reaction mixture was concentrated and the residue was purifiedby trituration with ether to afford V.

Yield: 2 g, crude: ¹H NMR (400 MHz, CDCl₃) δ 5.94-5.74 (m, 1H),5.30-5.02 (m, 2H), 3.15-3.03 (m, 2H), 2.86-2.73 (m, 2H).

Exemplary Procedure for the Preparation of methyl3-(cyclopropylmethoxy)-4-fluorobenzoate (VI)

To a stirred solution of 3-(cyclopropylmethoxy)-4-fluorobenzoic acid(3.00 g, 14.28 mmol) in MeOH (100 mL) was added conc.H₂SO₄ (1 mL) dropwise and the reaction mixture was heated at 65° C. for 8 h. The progressof the reaction was monitored by TLC. After completion of the reaction,the reaction mixture was neutralized with aqueous NaHCO₃ and evaporatedunder reduced pressure. The residue was diluted with water and extractedwith EtOAc. The combined organic layers were washed with brine, driedover anhydrous Na₂SO₄ and concentrated under reduced pressure. Theresidue was purified by column chromatography using 20% EtOAc/hexane toafford VI.

Yield: 2.3 g, 71.87%; ¹H NMR (400 MHz, DMSO-d₆) δ 7.65-7.53 (m, 2H),7.41-7.24 (m, 1H), 3.96 (d, J=7.0 Hz, 2H), 3.85 (s, 2H), 1.28-1.25 (m,1H), 0.58-0.56 (m, 2H), 0.36-0.33 (m, 2H).

Exemplary Procedure for the Preparation of(3-(cyclopropylmethoxy)-4-fluorophenyl) methanol (VII)

To a stirred solution of VI (2.30 g, 10.26 mmol) in dry DCM (20 mL), wasadded LiBH₄ (IM soln. in DCM, 20 mL, 20.53 mmol) drop wise at 0° C. andthe reaction mixture was heated at 80° C. for 12 h. The progress of thereaction was monitored by TLC. After completion of the reaction, thereaction mixture was quenched with water and extracted with EtOAc. Thecombined organic layers were washed with brine, dried over anhydrousNa₂SO₄ and concentrated under reduced pressure to afford VII.

Yield: 2 g, crude; ¹H NMR (400 MHz, DMSO-d₆) δ 7.36-7.00 (m, 2H),6.87-6.85 (m, 1H), 5.18 (t, J=5.7 Hz, 1H), 4.44 (d, J=5.7 Hz, 2H),4.08-3.75 (m, 2H), 1.32-1.13 (m, 1H), 0.64-0.49 (m, 2H), 0.35-0.32 (m,2H).

Exemplary Procedure for the Preparation of3-(cyclopropylmethoxy)-4-fluorobenzaldehyde (VIII)

To a stirred solution of VII (2.00 g, 10.20 mmol) in dry DCM (20 mL),PCC (4.38 g, 20.40 mmol) was added and warmed to room temperature for 5h. The progress of the reaction was monitored by TLC. After completionof the reaction, the reaction mixture was filtered through celite andthe filtrate was concentrated under reduced pressure to afford VIII.

Yield: 2.00 g, crude; ¹H NMR (400 MHz, DMSO-d6) δ 9.92 (s, 1H),7.71-7.52 (m, 2H), 7.52-7.06 (m, 1H), 3.99 (d, J=7.1 Hz, 2H), 1.34-1.16(m, 1H), 0.68-0.51 (m, 2H), 0.38-0.36 (m, 2H).

Exemplary Procedure for the Preparation of (Z)—N-(tert-butyl(1-oxidanyl)-l3-sulfanyl)-1-(3-(cyclopropylmethoxy)-4-fluorophenyl)methanimine (IX)

To a stirred solution of VIII (2.00 g, 10.25 mmol) andtert-butyl-3-sulfanamine (1.2 g, 10.25 mmol) in dry toluene (20 mL),Ti(O^(i)Pr)₄ (5.82 g, 20.50 mmol) was added and heated at 90° C. for 12h. The progress of the reaction was monitored by TLC. After completionof the reaction, the reaction mixture was filtered through celite andthe filtrate was diluted with water and extracted with EtOAc. Thecombined organic layers were washed with brine, dried over anhydrousNa₂SO₄ and evaporated under reduced pressure. The residue was purifiedby column chromatography using 10% EtOAc/hexane to afford IX.

Yield: 2.3 g, 75.65%; NMR: ¹H NMR (400 MHz, DMSO-d₆) δ 8.50 (s, 1H),7.69 (dd, J=8.4, 2.0 Hz, 1H), 7.57-7.55 (m, 1H), 7.38 (dd, J=11.2, 8.4Hz, 1H), 5.75 (s, 1H), 4.01-3.91 (m, 2H), 1.18 (s, 9H), 0.64-0.52 (m,2H), 0.40-0.28 (m, 2H).

Exemplary Procedure for the Preparation of(R)-1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethan-1-amine (X)

To a stirred solution of IX (2.30 g, 38.72 mmol) in dry THF (40 mL), wasadded a solution of CH₃MgBr (2M soln. in THF, 7.7 mL, 77.44 mmol) dropwise at 0° C. and the reaction mixture was stirred at room temperaturefor 2 h. The progress of the reaction was monitored by TLC. Aftercompletion of the reaction, the reaction mixture was quenched with waterand extracted with EtOAc. The combined organic layers were washed withbrine, dried over anhydrous Na₂SO₄ and evaporated under reducedpressure. A solution of HCl in dioxane (4 mL) was added to the residueand the reaction mixture was stirred at room temperature for 12 h. Aftercompletion of the reaction, the reaction mixture was concentrated andthe residue was purified by trituration with ether to afford X.

Yield: 2 g, NMR: ¹H NMR (400 MHz, DMSO-d₆) δ 8.64 (s, 2H), 7.48 (dd,J=8.3, 2.2 Hz, 1H), 7.22 (dd, J=11.4, 8.3 Hz, 1H), 7.07-7.04 (m, 1H),4.34 (p, J=5.9 Hz, 1H), 3.93 (d, J=7.1 Hz, 2H), 1.50 (d, J=6.7 Hz, 3H),1.30-1.24 (m, 1H), 0.60-0.58 (m, 2H), 0.40-0.27 (m, 2H).

Exemplary Procedure for the Preparation of(R)—N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl) ethyl)but-3-ene-1-sulfonamide (XI)

To a stirred solution of X (0.2 g, 0.93 mmol) in dry DCM (5 mL), Et₃N(0.241 g, 2.39 mmol) was added and stirred at rt for 10 min. After thatV (0.176 g, 1.14 mmol) in DCM (5 mL) was added drop wise and stirred atrt for 2 h. The progress of the reaction was monitored by TLC. Aftercompletion of the reaction, the reaction mixture was quenched with waterand extracted with DCM. The combined organic layer were washed withbrine, dried over anhydrous Na₂SO₄ and concentrated under reducedpressure. The crude product was purified by column chromatography using3% MeOH in DCM to afford XI.

Yield: 0.15 g, 48%.

Production Example 3. Synthesis of (R,E)-N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

To a stirred solution of compound XI (0.292 g, 0.89 mmol) and compoundIII (0.125 g, 0.89 mmol) in DCM (2 mL), Grubb's catalyst II^(nd)generation (0.015 g, 0.017 mmol) was added and the reaction mixture wasstirred at room temperature for 24 h. The progress of the reaction wasmonitored by TLC. After completion of the reaction, the reaction mixturewas concentrated under reduced pressure. The residue was purified bycolumn chromatography using 60% EtOAc/hexane to afford(R,E)-N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide.

Yield: 0.07 g, 17%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 7.70 (d,J=8.9 Hz, 1H), 7.19-7.15 (m, 2H), 6.95-6.89 (m, 1H), 5.49-5.45 (m, 1H),5.42-5.23 (m, 1H), 4.47-4.35 (m, 1H), 3.92-3.70 (m, 2H), 3.85 (s, 2H),3.79 (d, J=7.5 Hz, 2H), 2.89-2.81 (m, 2H), 2.75-2.70 (m, 2H), 2.31-2.17(m, 2H), 1.43-1.14 (m, 4H), 0.60-0.57 (m, 2H), 0.38-0.29 (m, 2H); ESI-MS(m/z): Calculated for: C₂₀H₂₆FN₃O₅S: 439.50; observed mass; 456.97(M+H₂O); HPLC purity: 99.5%; R_(t): 5.8

Production Example 8. Synthesis of (R,E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-isobutoxyphenyl)ethyl)pent-3-ene-1-sulfonamide

The title compound was prepared using 1-allylimidazolidine-2,4-dione and(R)—N-(1-(4-fluoro-3-isobutoxyphenyl)ethyl)but-3-ene-1-sulfonamide and1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.075 g, 14%; ¹H NMR (400 MHz, DMSO-d6) δ 10.75 (s, 1H), 7.73(dd, J=8.9, 1.7 Hz, 1H), 7.26-7.08 (m, 2H), 6.93-6.89 (m, 1H), 5.53-5.41(m, 1H), 5.34-5.23 (m, 1H), 4.48-4.35 (m, 1H), 3.87-3.70 (m, 6H),2.90-2.87 (m, 1H), 2.63-2.59 (m, 1H), 2.23-2.19 (m, 2H), 2.06-2.01 (m,1H), 1.37 (d, J=7.0 Hz, 3H), 0.99 (d, J=6.9, Hz, 6H); ESI-MS (m/z):Calculated for: C₂₀H₂₈FN₃O₅S: 441.52; observed mass; 464.10 (M+Na); HPLCpurity: 99.3%; R_(t); 8.2

Production Example 26. Synthesis of (R,E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-(neopentyloxy)phenyl)ethyl)pent-3-ene-1-sulfonamide

The title compound was prepared using(R)—N-(1-(4-fluoro-3-(neopentyloxy)phenyl)ethyl)but-3-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.05 g, 20%; ¹H NMR (400 MHz, DMSO-d⁶) δ 10.7 (s, 1H), 7.73 (d,J=8.6 Hz, 1H), 7.22 (d, J=8.1 Hz, 1H), 7.16-7.12 (m, 1H), 6.95-6.87 (m,1H), 5.49-5.46 (m, 1H), 5.30-5.27 (m, 1H), 4.41 (t, J=8.2 Hz, 1H), 3.79(s, 2H), 3.75-3.64 (m, 4H), 2.90-2.87 (m, 1H), 2.41-2.22 (m, 1H),2.24-2.20 (m, 2H), 1.40-1.31 (m, 3H), 1.01 (s, 9H); ESI-MS (m/z):Calculated for: C₂₁H₃₀FN₃O₅S: 455.55; observed mass: 473.20 (M+H₂O);HPLC purity: 97.1%; R_(t); 8.6

Production Example 28. Synthesis of (R,E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-(2-hydroxy-2-methylpropoxy)phenyl)ethyl)pent-3-ene-1-sulfonamide

The title compound was prepared using(R)—N-(1-(4-fluoro-3-(2-hydroxy-2-methylpropoxy)phenyl)ethyl)but-3-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.11 g, 36%; ¹H NMR (400 MHz, DMSO-d⁶) δ 10.75 (s, 1H), 7.74 (d,J=8.8 Hz, 1H), 7.23 (dd, J=8.3, 2.1 Hz, 1H), 7.13 (dd, J=11.4, 8.3 Hz,1H), 6.93-6.90 (m, 1H), 5.47-5.42 (m, 1H), 5.29-5.20 (m, 1H), 4.69 (s,1H), 4.48-4.35 (m, 1H), 3.82-3.69 (m, 6H), 2.90-2.88 (m, 1H), 2.66-6.32(m, 1H), 2.33-2.12 (m, 2H), 1.37 (d, J=6.9 Hz, 3H), 1.21 (s, 6H); ESI-MS(m/z): Calculated for: C₂₀H₂₈FN₃)₆S: 457.52; observed mass; 456.10(M−H); HPLC purity: 99.8%; R_(t); 6.8

Production Example 29. Synthesis of (R,E)-N-(1-(3-(cyclopropylmethoxy)phenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

The title compound was prepared using(R)—N-(1-(3-(cyclopropylmethoxy)phenyl)ethyl)but-3-ene-1-sulfonamidetheand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.09 g, 16.5%; ¹H NMR (400 MHz, DMSO-d6) δ 10.7 (s, 1H), 7.74 (d,J=8.8 Hz, 1H), 7.21 (t, J=7.8 Hz, 1H), 7.01-6.88 (m, 2H), 6.79-6.74 (m,1H), 5.41-5.38 (m, 1H), 5.32-5.20 (m, 1H), 4.46-4.32 (m, 1H), 3.87-3.64(m, 6H), 2.88-2.83 (m, 1H), 2.58-2.52 (m, 1H), 2.25-2.20 (m, 2H),1.41-1.31 (m, 3H), 1.29-1.13 (m, 1H), 0.61-0.50 (m, 2H), 0.38-0.24 (m,2H); ESI-MS (m/z): Calculated for: C₂₀H₂₇N₃O₅S: 421.51; observed mass:422.10 (M+H); HPLC purity: 94.1%; R_(t); 7.7

Production Example 57. Synthesis of [(R,E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(3-isobutoxyphenyl)ethyl]pent-3-ene-1-sulfonamide

The title compound was prepared using(R)—N-(1-(3-isobutoxyphenyl)ethyl)but-3-ene-1-sulfonamide and1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.16 g, 25%; ¹H NMR (400 MHz, DMSO-d⁶) δ 10.7 (s, 1H), 7.74 (d,J=8.6 Hz, 1H), 7.21 (t, J=7.9 Hz, 1H), 6.99 (s, 1H), 6.91 (d, J=7.5 Hz,1H), 6.78 (dd, J=8.5, 2.6 Hz, 1H), 5.45-5.40 (m, 1H), 5.28-5.23 (m, 1H),4.39-4.30 (m, 1H), 3.74-3.70 (m, 6H), 2.88-2.83 (m, 1H), 2.59-2.54 (m,1H), 2.25-2.20 (m, 2H), 2.04-1.99 (m, 1H), 1.37 (d, J=6.8 Hz, 3H), 0.98(d, J=6.7 Hz, 6H); ESI-MS (m/z): Calculated for: C₂₀H₂₉N₃O₅S: 423.53;observed mass; 422.20 (M−1); HPLC purity: 97.3%; R_(t); 7.9

Production Example 67. Synthesis of (R,E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-(2-hydroxy-2-methylpropoxy)phenyl)ethyl)pent-3-ene-1-sulfonamide

The title compound was prepared using(R)—N-(1-(4-fluoro-3-(2-hydroxy-2-methylpropoxy)phenyl)ethyl)but-3-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.11 g, 36%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 7.74 (d,J=8.8 Hz, 1H), 7.23 (dd, J=8.3, 2.1 Hz, 1H), 7.13 (dd, J=11.4, 8.3 Hz,1H), 6.93-6.90 (m, 1H), 5.47-5.40 (m, 1H), 5.29-5.52 (m, 1H), 4.69 (s,1H), 4.48-4.35 (m, 1H), 3.82-3.69 (m, 6H), 2.90-2.88 (m, 1H), 2.66-6.32(m, 1H), 2.33-2.12 (m, 2H), 1.37 (d, J=6.9 Hz, 3H), 1.21 (s, 6H); ESI-MS(m/z): Calculated for: C₂₀H₂₈FN₃O₆S: 457.52; observed mass; 456.10(M−H); HPLC purity: 99.8%; R_(t): 6.8

Production Example 43. Synthesis of (R,E)-N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)propyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

The title compound was prepared using(R)—N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)propyl)but-3-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.108 g, 12.5%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 7.70(d, J=9.4 Hz, 1H), 7.22-7.08 (m, 2H), 6.90 (d, J=5.0 Hz, 1H), 5.47-5.29(m, 1H), 5.23-5.20 (m, 1H), 4.11-4.08 (m, 1H), 3.92-3.84 (m, 2H), 3.78(s, 2H), 3.71 (d, J=5.7 Hz, 2H), 2.83-2.79 (m, 1H), 2.28-2.07 (m, 2H),1.66-1.62 (m, 2H), 1.25-1.20 (m, 3H), 0.85-0.77 (m, 3H), 0.58-0.50 (m,2H), 0.32-0.30 (m, 2H); ESI-MS (m/z): Calculated for: C₂₁H₂₈FN₃O₅S:453.53 observed mass: 454.10 (M+1); HPLC purity: 99.9%; R_(t); 8.0

Production Example 95. Synthesis of (R,E)-N-(1-(3-(cyclopropylmethoxy)phenyl)propyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

The title compound was prepared using(R)—N-(1-(3-(cyclopropylmethoxy)phenyl)propyl)but-3-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.095 g, 17.6%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 7.73(d, J=9.1 Hz, 1H), 7.20 (t, J=7.9 Hz, 1H), 6.96-6.92 (m, 1H), 6.88 (d,J=7.5 Hz, 1H), 6.77 (dd, J=8.2, 2.4 Hz, 1H), 5.35-5.30 (m, 1H),5.20-5.17 (m, 1H), 4.08-4.00 (m, 1H), 3.89-3.62 (m, 6H), 2.81-2.78 (m,1H), 2.45-2.42 (m, 1H), 2.21-2.18 (m, 2H), 1.76-1.55 (m, 2H), 1.23-1.20(m, 1H), 0.82 (t, J=7.3 Hz, 3H), 0.61-0.50 (m, 2H), 0.38-0.26 (m, 2H);ESI-MS (m/z): Calculated for: C₂₁H₂₉N₃O₅S: 435.54 observed mass; 434.20(M−H); HPLC purity: 99.1%; R_(t); 7.7

Production Example 1. Synthesis of(R)—N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

To a stirred solution of(R,E)-N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide(0.06 g, 0.13 mmol) in MeOH (4 mL), Rh/Al₂O₃ (6 mg) was added andstirred under hydrogen atmosphere (balloon pressure) at room temperaturefor 16 h. The progress of the reaction was monitored by TLC. Aftercompletion of the reaction, the reaction mixture was filtered throughcelite and filtrate was evaporated under reduced pressure. The residuewas purified by column chromatography using 50% EtOAc/hexane to afford(R)—N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide.

Yield: 0.03 g, 50%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 7.66 (d,J=8.8 Hz, 1H), 7.23-7.09 (m, 2H), 6.95-6.89 (m, 1H), 4.46-4.33 (m, 1H),3.89-3.84 (m, 4H), 3.13 (t, J=7.0 Hz, 2H), 2.80-2.76 (m, 1H), 2.56-2.32(m, 1H), 1.57-1.00 (m, 10H), 0.60-0.57 (m, 2H), 0.33-0.30 (m, 2H);ESI-MS (m/z): Calculated for: C₂₀H₂₈FN₃O₅S: 441.52 observed mass; 464.15(M+Na); HPLC purity: 95.9%; R_(t): 7.8

Production Example 20. Synthesis of5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)pentane-1-sulfonamide

The title compound was prepared using(E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.045 g, 56%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.17 (s,1H), 7.24 (d, J=8.1 Hz, 1H), 7.12 (dd, J=11.3, 8.4 Hz, 1H), 6.96-6.88(m, 1H), 3.89-3.77 (m, 4H), 3.12 (t, J=7.1 Hz, 2H), 2.54 (d, J=7.3 Hz,2H), 2.06-2.02 (m, 1H), 1.43-1.40 (m, 2H), 1.35-1.14 (m, 4H), 1.11-0.95(m, 10H); ESI-MS (m/z): Calculated for: C₂₁H₃₀FN₃O₅S: 455.55; observedmass: 456.20 (M+H); HPLC purity: 99.4%; R_(t); 8.2

Production Example 22. Synthesis of(R)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-(2-hydroxy-2-methylpropoxy)phenyl)ethyl)pentane-1-sulfonamide

The title compound was prepared using (R,E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-(2-hydroxy-2-methylpropoxy)phenyl)ethyl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.09 g, 93%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 7.69 (d,J=8.7 Hz, 1H), 7.23 (dd, J=8.3, 2.0 Hz, 1H), 7.19-7.09 (m, 1H),6.93-6.90 (m, 1H), 4.69 (s, 1H), 4.47-4.34 (m, 1H), 3.87 (s, 2H), 3.77(s, 2H), 3.13 (t, J=7.1 Hz, 2H), 2.81-2.78 (m, 1H), 2.61-2.50 (m, 1H),1.60-1.40 (m, 2H), 1.34-1.30 (m, 5H), 1.30 (s, 6H), 1.10-1.00 (m, 2H);ESI-MS (m/z): Calculated for: C₂₀H₃₀FN₃O₆S: 459.53; observed mass:482.20 (M+Na); HPLC purity: 93.3%; R_(t); 6.9

Production Example 63. Synthesis of(R)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(3-isobutoxyphenyl)ethyl)pentane-1-sulfonamide

The title compound was prepared using [(R,E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(3-isobutoxyphenyl)ethyl]pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.1 g, 77%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 7.67 (d,J=8.7 Hz, 1H), 7.22 (t, J=7.9 Hz, 1H), 6.99 (s, 1H), 6.91 (d, J=7.6 Hz,1H), 6.80 (d, J=8.2 Hz, 1H), 4.38-4.30 (m, 1H), 3.86 (s, 2H), 3.72 (d,J=6.4 Hz, 2H), 3.22-3.08 (m, 2H), 2.78-2.74 (m, 1H), 2.58-2.44 (m, 1H),2.04-1.99 (m, 1H), 1.58-1.21 (m, 6H), 1.19-0.91 (m, 9H); ESI-MS (m/z):Calculated for: C₂₀H₃₁N₃O₅S: 425.54; observed mass: 424.15 (M−H); HPLCpurity: 96.8%; R_(t); 7.9

Production Example 64. Synthesis of(R)—N-(1-(3-(cyclopropylmethoxy)phenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

The title compound was prepared using(R,E)-N-(1-(3-(cyclopropylmethoxy)phenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.04 g, 61.3%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 7.68 (d,J=8.8 Hz, 1H), 7.21 (t, J=7.9 Hz, 1H), 7.00-6.88 (m, 2H), 6.82-6.74 (m,1H), 4.48-4.31 (m, 1H), 3.87 (s, 2H), 3.79 (d, J=7.0 Hz, 2H), 3.12 (t,J=7.0 Hz, 2H), 2.78-2.74 (m, 1H), 2.58-2.42 (m, 1H), 1.58-0.96 (m, 10H),0.61-0.50 (m, 2H), 0.38-0.27 (m, 2H); ESI-MS (m/z): Calculated for:C₂₀H₂₉N₃O₅S: 423.53; observed mass; 422.10 (M−H); HPLC purity: 99.9%;R_(t); 7.5

Production Example 80. Synthesis of(R)—N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)propyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

The title compound was prepared using(R,E)-N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)propyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.082 g, 93%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 7.64 (d,J=9.2 Hz, 1H), 7.22-7.09 (m, 2H), 6.90-6.88 (m, 1H), 4.10 (q, J=7.9 Hz,1H), 3.92-3.81 (m, 4H), 3.16-3.04 (m, 2H), 2.74-2.70 (m, 1H), 2.48-244(m, 1H), 1.67-1.64 (m, 2H), 1.52-1.42 (m, 1H), 1.41-1.34 (m, 1H),1.33-1.19 (m, 3H), 1.09-1.05 (m, 2H), 0.81 (t, J=7.3 Hz, 3H), 0.6-0.57(m, 2H), 0.39-0.27 (m, 2H); ESI-MS (m/z): Calculated for: C₂₁H₃₀FN₃O₅S:455.55; observed mass: 456.10 (M+H); HPLC purity: 98.4%; R_(t); 8.0

Production Example 96. Synthesis of(R)—N-(1-(3-(cyclopropylmethoxy)phenyl)propyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

The title compound was prepared using(R,E)-N-(1-(3-(cyclopropylmethoxy)phenyl)propyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.082 g, 93%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 7.66 (d,J=9.3 Hz, 1H), 7.21 (t, J=7.9 Hz, 1H), 6.99-6.93 (m, 1H), 6.88 (d, J=7.6Hz, 1H), 6.78 (dd, J=8.1, 2.6 Hz, 1H), 4.07 (q, J=7.9 Hz, 1H), 3.85 (s,1H), 3.79 (d, J=7.0 Hz, 2H), 3.14-3.05 (m, 2H), 2.72-2.69 (m, 1H),2.39-2.35 (m, 1H), 1.69-1.64 (m, 2H), 1.43-1.15 (m, 6H), 1.09-1.04 (m,1H), 0.96-0.92 (m, 1H), 0.82 (t, J=7.2 Hz, 3H), 0.61-0.50 (m, 2H),0.33-0.30 (m, 2H); ESI-MS (m/z): Calculated for: C₂₁H₃₁N₃O₅S: 437.56;observed mass: 436.15 (M−H); HPLC purity: 93.4%; R_(t); 7.8

Production Example 21: Synthesis of (R,E)-N-(1-(3-(2,2-difluoroethoxy)-4-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

The title compound was prepared using(R)—N-(1-(3-(2,2-difluoroethoxy)-4-fluorophenyl)ethyl)but-3-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.051 g, 13%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (s, 1H), 7.72 (d,J=8.8 Hz, 1H), 7.33-7.16 (m, 2H), 7.025-7.01 (m, 1H), 6.42 (tt, J=56.0,3.6 Hz, 1H), 5.55-5.36 (m, 1H), 5.34-5.29 (m, 1H), 4.50-4.27 (m, 3H),3.87-3.68 (m, 4H), 2.95-2.89 (m, 1H), 2.73-2.69 (m, 1H), 2.24-2.22 (m,2H), 1.38 (d, J=6.8 Hz, 3H); ESI-MS (m/z): Calculated for:C₁₈H₂₂F₃N₃O₅S: 449.45: observed mass; 472.90 (M+Na); HPLC purity: 95.5%;R_(t); 7.4

Production Example 23 Synthesis of(R)—N-(1-(3-(2,2-difluoroethoxy)-4-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

The title compound was prepared using (R,E)-N-(1-(3-(2,2-difluoroethoxy)-4-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.03 g, 86%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 7.66 (d,J=8.8 Hz, 1H), 7.33-7.16 (m, 2H), 7.05-7.01 (m, 1H), 6.42 (tt, J=56.0,3.6 Hz, 1H), 4.4-4.37 (m, 3H), 3.87 (s, 2H), 3.13 (t, J=7.0 Hz, 2H),2.82-2.79 (m, 1H), 2.63-2.60 (m, 1H), 1.60-1.27 (m, 9H); ESI-MS (m/z):Calculated for: C₁₈H₂₄F₃N₃O₅S: 451.46; observed mass: 469.10 (M+H₂O);HPLC purity: 97.9%; R_(t); 7.5

Exemplary Procedure for the Preparation of3-(cyclopropylmethoxy)-4-fluorobenzonitrile (XII)

To a stirred solution of 4-fluoro-3-hydroxybenzonitrile (10.0 g, 78.74mmol) in dry DMF (100 mL), K₂CO₃ (21.73 g, 157.4 mmol) was addedfollowed by addition of cyclopropyl methyl bromide (12.85 g, 94.48mmol). The reaction mixture was heated at 90° C. for 4 h. The progressof the reaction was monitored by TLC. After completion of the reaction,the reaction mixture was quenched with cold water and precipitated solidwas filtered, washed with pentane to afford XII

Yield: 12 g, 94%; ¹H NMR (400 MHz, CDCl₃) δ 7.28-7.11 (m, 3H), 3.90 (d,J=7.1 Hz, 2H), 1.32-1.29 (m, 1H), 0.76-0.63 (m, 2H), 0.45-0.32 (m, 2H).

Exemplary Procedure for the Preparation of1-(3-(cyclopropylmethoxy)-4-fluorophenyl)cyclopropan-1-amine (XIII)

To a stirred solution of XII (7.8 g, 40.83 mmol) in dry THF (40 mL),Ti(O^(i)Pr)₄ (12.75 g, 44.92 mmol) was added at −78° C. CH₃CH₂MgBr (3Msoln. in Et₂O, 29 mL, 89.82 mmol) was added drop wise under nitrogenatmosphere and the reaction mixture was stirred at room temperature atfor 1 h. BF₃.OEt₂ (5.68 g, 80.0 mmol) was added drop wise and stirred atroom temperature for 1.5 h. The progress of the reaction was monitoredby TLC. After completion of the reaction, the reaction mixture wasdiluted with 1N HCl and stirred for 10 min. After that reaction mixturewas neutralized with aqueous NaOH and extracted with Et₂O. The combinedorganic layers were washed with brine, dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography using 30% EtOAC/hexane to afford XIII.

Yield: 4.1 g, 47%. NMR: 1H NMR (400 MHz, DMSO-d₆) δ 4.04 (d, J=7.1 Hz,2H), 2.54 (d, J=9.9 Hz, 2H), 2.48-2.27 (m, 5H), 1.90-1.72 (m, 2H),1.17-1.10 (m, J=7.0 Hz, 2H).

Exemplary Procedure for the Preparation ofN-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)cyclopropyl)but-3-ene-1-sulfonamide(XIV)

To a stirred solution of XIII (0.1 g, 0.44 mmol) in dry DCM (4 mL), Et₃N(0.08 mL, 0.57 mmol) was added and stirred at room temperature for 10min. After that V (0.083 g, 0.53 mmol) in DCM (4 mL) was added drop wiseand stirred at room temperature for 4 h. The progress of the reactionwas monitored by TLC. After completion of the reaction, the reactionmixture was quenched with water and extracted with EtOAc. The combinedorganic layer were washed with brine, dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The crude product was purified bycolumn chromatography using 30% EtOAc/hexane in DCM to afford XIV.

Yield: 0.043 g, 30%; NMR: ¹H NMR (400 MHz, CDCl₃) δ 7.11-6.92 (m, 3H),5.60-5.55 (m, 1H), 5.10 (s, 1H), 5.04-4.89 (m, 2H), 3.89 (d, J=6.9 Hz,2H), 2.79-2.63 (m, 2H), 2.36-2.25 (m, 2H), 1.56 (d, J=0.9 Hz, 1H),1.44-1.24 (m, 4H), 1.24-1.10 (m, 2H), 0.72-0.60 (m, 2H). ESI-MS (m/z);342.10 (M+H);

Production Example 8: Synthesis of(E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)pent-3-ene-1-sulfonamide

The title compound was prepared usingN-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)but-3-ene-1-sulfonamide and1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.1 g, 15.2%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.17 (s,1H), 7.24 (d, J=8.1 Hz, 1H), 7.12 (dd, J=11.3, 8.4 Hz, 1H), 6.96-6.88(m, 1H), 5.53-5.41 (m, 1H), 5.34-5.23 (m, 1H), 3.89-3.77 (m, 4H),3.86-3.76 (m, 2H), 2.66-2.52 (m, 2H), 2.15 (m, 1H), 2.10-2.00 (m, 1H),1.23 (s, 4H), 1.12-0.90 (m, 7H); ESI-MS (m/z): Calculated for:C₂₁H₂₈FN₃O₅S: 453.53; observed mass: 454.1 (M+H); HPLC purity: 98.5%;R_(t); 8.2

Production Example 10: Synthesis of(E)-N-(1-(3-(cyclobutylmethoxy)-4-fluorophenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

The title compound was prepared usingN-(1-(3-(cyclobutylmethoxy)-4-fluorophenyl)cyclopropyl)but-3-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.1 g, 15.22%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.22 (s,1H), 7.24 (dd, J=8.3, 2.2 Hz, 1H), 7.22-7.05 (m, 1H), 6.95-6.90 (m, 1H),5.45-5.39 (m, 1H), 5.36-5.18 (m, 1H), 4.01 (d, J=6.7 Hz, 2H), 3.89 (s,2H), 3.79-3.69 (m, 1H), 2.78-2.72 (m, 1H), 2.66-2.57 (m, 2H), 2.32-1.75(m, 8H), 1.29-1.05 (m, 5H); ESI-MS (m/z): Calculated for: C₂₂H₂₈FN₃O₅S:465.54 observed mass; 466.20 (M+H); HPLC purity: 91.4%; R_(t); 8.4

Production Example 11: Synthesis of(E)-N-(1-(3-(2,2-difluoroethoxy)-4-fluorophenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

The title compound was prepared usingN-(1-(3-(2,2-difluoroethoxy)-4-fluorophenyl)cyclopropyl)but-3-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.03 g, 7.59%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.20 (s,1H), 7.25-7.13 (m, 2H), 7.05-7.02 (m, 1H), 6.20 (tt, J=56.0, 3.6 Hz,1H), 5.49-5.21 (m, 2H), 4.37 (dt, J=14.6, 3.6 Hz, 2H), 3.86 (s, 2H),3.72 (d, J=5.9 Hz, 2H), 2.67 (dd, J=8.8, 6.5 Hz, 2H), 2.17 (q, J=7.1 Hz,2H), 1.30-1.08 (m, 4H); ESI-MS (m/z): Calculated for: C₁₉H₂₂F₃N₃O₅S:461.46; observed mass: 462.05 (M+H); HPLC purity: 98.0%; R_(t); 7.4

Production Example 12: Synthesis of(E)-N-(1-(3-(cyclopentylmethoxy)-4-fluorophenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

The title compound was prepared usingN-(1-(3-(cyclopentylmethoxy)-4-fluorophenyl)cyclopropyl)but-3-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.08 g, 12.3%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.23 (d,J=6.1 Hz, 1H), 7.29-7.16 (m, 1H), 7.16-7.06 (m, 1H), 6.96-6.88 (m, 1H),5.41-5.38 (m, 1H), 5.25-5.20 (m, 1H), 3.90 (d, J=6.8 Hz, 2H), 3.78 (s,2H), 3.72 (d, J=6.0 Hz, 2H), 3.36-3.20 (m, 1H), 2.65-2.56 (m, 2H),2.35-2.23 (m, 1H), 2.15-2.00 (m, 1H), 1.79-1.75 (m, 2H), 1.59-1.55 (m,4H), 1.39-1.04 (m, 6H); ESI-MS (m/z): Calculated for: C₂₃H₃₀FN₃O₅S:479.57; observed mass: 480.20 (M+H); HPLC purity: 98.3%; R_(t); 8.6

Production Example 13: Synthesis of(E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-(neopentyloxy)phenyl) cyclopropyl) pent-3-ene-1-sulfonamide

The title compound was prepared usingN-(1-(4-fluoro-3-(neopentyloxy)phenyl)cyclopropyl)but-3-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.045 g, 15%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (s, 1H), 8.22 (s,1H), 7.25 (dd, J=8.3, 2.2 Hz, 1H), 7.11 (dd, J=11.4, 8.5 Hz, 1H), 5.41(d, J=15.4 Hz, 1H), 5.29-5.19 (m, 1H), 3.80-3.66 (m, 6H), 2.66-2.57 (m,2H), 2.16-2.13 (m, 2H), 1.23 (d, J=4.7 Hz, 3H), 1.09 (t, J=3.6 Hz, 2H),1.01 (s, 9H); ESI-MS (m/z): Calculated for: C₂₂H₃₀FN₃O₅S: 467.56;observed mass: 468.05 (M+H); HPLC purity: 98.7%; R_(t); 8.5

Production Example 14: Synthesis of(E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-(2-hydroxy-2-methylpropoxy)phenyl)cyclopropyl)pent-3-ene-1-sulfonamide

The title compound was prepared usingN-(1-(4-fluoro-3-(2-hydroxy-2-methylpropoxy)phenyl)cyclopropyl)but-3-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.035 g, 5.32%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (s, 1H), 8.23(s, 1H), 7.25-7.08 (m, 2H), 6.96-6.92 (m, 1H), 5.37-5.31 (m, 1H),5.23-5.19 (m, 1H), 4.64 (s, 1H), 3.88 (d, J=7.1 Hz, 4H), 3.51 (d, J=6.0Hz, 2H), 2.61-2.52 (m, 2H), 2.15-2.12 (m, 2H), 1.38-1.03 (m, 8H),0.90-0.76 (m, 2H); ESI-MS (m/z): Calculated for: C₂₁H₂₈FN₃O₆S: 469.53;observed mass: 491.75 (M+Na); HPLC purity: 97.5%; R_(t); 6.8

Production Example 60: Synthesis of(E)-N-(1-(3-(2,2-difluoroethoxy)phenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

The title compound was prepared usingN-(1-(3-(2,2-difluoroethoxy)phenyl)cyclopropyl)but-3-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.076 g, 19%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (s, 1H), 8.24 (s,1H), 7.24-7.20 (m, 1H), 7.07-6.96 (m, 2H), 6.88-6.74 (m, 1H), 6.34 (tt,J=56.0, 3.7 Hz, 1H), 5.45-5.33 (m, 1H), 5.32-5.20 (m, 1H), 4.28 (dt,J=14.8, 3.6 Hz, 2H), 3.89-3.75 (m, 2H), 3.72 (d, J=5.7 Hz, 2H),2.65-2.61 (m, 2H), 2.18-2.15 (m, 2H), 1.30-1.14 (m, 2H), 1.10-1.04 (m,2H); ESI-MS (m/z): Calculated for: C₁₉H₂₃F₂N₃O₅S: 443.47 observed mass;443.75 (M+H); HPLC purity: 98.0%; R_(t); 7.3

Production Example 57: Synthesis of(E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(3-isobutoxyphenyl)cyclopropyl)pent-3-ene-1-sulfonamide

The title compound was prepared usingN-(1-(3-isobutoxyphenyl)cyclopropyl)but-3-ene-1-sulfonamide and1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.06 g, 11%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (s, 1H), 8.25 (s,1H), 7.19 (t, J=8.0 Hz, 1H), 7.04 (s, 1H), 6.92 (d, J=7.5 Hz, 1H),6.80-6.72 (m, 1H), 5.39-5.35 (m, 1H), 5.28-5.16 (m, 1H), 3.79-3.67 (m,6H), 2.63-2.55 (m, 2H), 2.16-2.13 (m, 2H), 2.02-1.98 (m, 1H), 1.30-1.03(m, 4H), 0.97 (d, J=6.5 Hz, 6H); ESI-MS (m/z): Calculated for:C₂₁H₂₉N₃O₅S: 435.54 observed mass; 435.85 (M+H); HPLC purity: 93.4%;R_(t); 8.0

Production Example 58: Synthesis of(E)-N-(1-(3-(cyclopropylmethoxy)phenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

The title compound was prepared usingN-(1-(3-(cyclopropylmethoxy)phenyl)cyclopropyl)but-3-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.11 g, 16.3%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (s, 1H), 8.22 (s,1H), 7.17 (t, J=7.9 Hz, 1H), 6.99 (s, 1H), 6.92 (d, J=7.7 Hz, 1H), 6.73(dd, J=8.0, 2.6 Hz, 1H), 5.36-5.31 (m, 1H), 5.25-5.20 (m, 1H), 3.77-3.72(m, 4H), 3.70 (d, J=5.8 Hz, 2H), 2.59 (dd, J=9.8, 6.1 Hz, 2H), 2.15-2.12(m, 2H), 1.21-1.15 (m, 3H), 1.06-1.00 (m, 2H), 0.60-0.50 (m, 2H),0.30-0.26 (m, 2H); ESI-MS (m/z): Calculated for: C₂₁H₂₇N₃O₅S: 433.52;observed mass: 434.10 (M+H); HPLC purity: 99.5%; R_(t): 7.6

Production Example 5: Synthesis ofN-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

The title compound was prepared using(E)-N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.09 g, 90%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.17 (s,1H), 7.24 (d, J=8.1 Hz, 1H), 7.12 (dd, J=11.3, 8.4 Hz, 1H), 6.96-6.88(m, 1H), 3.89-3.77 (m, 4H), 3.12-3.10 (m, 2H), 2.56-2.53 (m, 2H),1.42-1.03 (m, 11H), 0.65-0.60 (m, 2H), 0.38-0.32 (d, J=5.0 Hz, 2H);ESI-MS (m/z): Calculated for: C₂₁H₂₈FN₃O₅S: 453.53; observed mass:454.10 (M+H); HPLC purity: 99.3%; R_(t); 7.9

Production Example 7: Synthesis of5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)pentane-1-sulfonamide

The title compound was prepared using(E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.045 g, 56%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.17 (s,1H), 7.24 (d, J=8.1 Hz, 1H), 7.12 (dd, J=11.3, 8.4 Hz, 1H), 6.96-6.88(m, 1H), 3.89-3.77 (m, 4H), 3.12 (t, J=7.1 Hz, 2H), 2.54-2.50 (m, 2H),2.06-2.02 (m, 1H), 1.43-1.40 (m, 2H), 1.35-1.14 (m, 4H), 1.11-0.95 (m,10H); ESI-MS (m/z): Calculated for: C₂₁H₃₀FN₃O₅S: 455.55; observed mass456.20 (M+H); HPLC purity: 99.4%; R_(t); 8.2

Production Example 9: Synthesis ofN-(1-(3-(cyclobutylmethoxy)-4-fluorophenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

The title compound was prepared using(E)-N-(1-(3-(cyclobutylmethoxy)-4-fluorophenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.06 g, 60%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.17 (s,1H), 7.24 (dd, J=8.3, 2.2 Hz, 1H), 7.12 (dd, J=11.4, 8.4 Hz, 1H),6.95-6.91 (m, 1H), 4.02 (d, J=6.7 Hz, 2H), 3.87 (s, 2H), 3.12 (t, J=7.1Hz, 2H), 2.78-2.71 (m, 1H), 2.54-2.51 (m, 2H), 2.10-2.05 (m, 2H),1.96-1.75 (m, 4H), 1.49-1.37 (m, 2H), 1.36-0.98 (m, 8H); ESI-MS (m/z):Calculated for: C₂₂H₃₀FN₃O₅S: 467.56; observed mass 468.10 (M+H); HPLCpurity: 99.2%; R_(t); 8.3

Production Example 35: Synthesis ofN-(1-(3-(cyclopentylmethoxy)-4-fluorophenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

The title compound was prepared using(E)-N-(1-(3-(cyclopentylmethoxy)-4-fluorophenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.025 g, 40.2%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.17(s, 1H), 7.25 (dd, J=8.5, 2.2 Hz, 1H), 7.12 (dd, J=11.4, 8.4 Hz, 1H),6.95-6.91 (m, 1H), 3.94-3.80 (m, 4H), 3.12 (t, J=7.0 Hz, 2H), 2.63-2.47(m, 2H), 2.35-2.30 (m, 1H), 1.84-1.71 (m, 2H), 1.68-0.97 (m, 16H);ESI-MS (m/z): Calculated for: C₂₃H₃₂FN₃O₅S: 481.58; observed mass;482.15 (M+H); HPLC purity: 95.8%; R_(t); 8.7

Production Example 36: Synthesis ofN-(1-(3-(2,2-difluoroethoxy)-4-fluorophenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

The title compound was prepared using(E)-N-(1-(3-(2,2-difluoroethoxy)-4-fluorophenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.05 g, 83.3%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.14 (s,1H), 7.19-7.15 (m, 2H), 7.05-7.00 (m, 1H), 6.40 (tt, J=56.0, 3.3 Hz,1H), 4.37 (dt, J=14.6, 3.5 Hz, 2H), 3.86 (s, 2H), 3.11 (t, J=7.1 Hz,2H), 2.62-2.53 (m, 2H), 1.43-1.40 (m, 2H), 1.36-1.00 (m, 8H); ESI-MS(m/z): Calculated for: C₁₉H₂₄F₃N₃O₅S: 463.47; observed mass; 464.07(M+H); HPLC purity: 97.1%; R_(t); 7.4

Production Example 37: Synthesis of5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-(neopentyloxy)phenyl)cyclopropyl)pentane-1-sulfonamide

The title compound was prepared using(E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-(neopentyloxy)phenyl) cyclopropyl) pent-3-ene-1-sulfonamide and Rh.Al₂O₃ in a mannersimilar to the method in Production Example 1 above.

Yield: 0.022 g, 73.3%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.18(s, 1H), 7.25 (dd, J=8.3, 2.2 Hz, 1H), 7.13 (dd, J=11.4, 8.4 Hz, 1H),6.96-6.91 (m, 1H), 3.86 (s, 2H), 3.69 (s, 2H), 3.12 (t, J=7.0 Hz, 2H),2.54-2.51 (m, 2H), 1.42-1.38 (m, 2H), 1.36-1.16 (m, 8H), 1.01 (s, 9H);ESI-MS (m/z): Calculated for: C₂₂H₃₂FN₃O₅S: 469.57 observed mass; 470.15(M+H); HPLC purity: 95.4%; R_(t); 8.7

Production Example 38: Synthesis of5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-(2-hydroxy-2-methylpropoxy)phenyl)cyclopropyl)pentane-1-sulfonamide

The title compound was prepared using(E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-(2-hydroxy-2-methylpropoxy)phenyl)cyclopropyl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.022 g, 73.3%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.16(s, 1H), 7.25 (d, J=8.4 Hz, 1H), 7.11 (t, J=10.0 Hz, 1H), 6.93-6.89 (m,1H), 4.67 (s, 1H), 3.86 (s, 2H), 3.76 (s, 2H), 3.11 (t, J=7.2 Hz, 2H),2.54 (dd, J=6.6, 4.1 Hz, 2H), 1.47-1.45 (m, 2H), 1.43-1.40 (m, 2H),1.33-1.14 (m, 8H), 1.05-0.98 (m, 4H); ESI-MS (m/z): Calculated for:C₂₁H₃₀F₃O₆S: 471.54; observed mass; 470 (M−H); HPLC purity: 98.1%;R_(t); 7.0

Production Example 75: Synthesis of (R,E)-N-(1-(3-(cyclopropylmethoxy)phenyl)propyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

The title compound was prepared using(R)—N-(1-(3-(cyclopropylmethoxy)phenyl)propyl)but-3-ene-1-sulfonamide ina manner similar to the method in Production Example 1 above.

Yield: 0.095 g, 17.6%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (s, 1H), 7.73(d, J=9.1 Hz, 1H), 7.20 (t, J=7.9 Hz, 1H), 6.96 (t, J=2.0 Hz, 1H), 6.88(d, J=7.5 Hz, 1H), 6.77 (dd, J=8.2, 2.4 Hz, 1H), 5.37-5.32 (m, 1H),5.20-5.17 (m, 1H), 4.08-4.02 (m, 1H), 3.89-3.62 (m, 6H), 2.81-2.78 (m,1H), 2.45-2.42 (m, 1H), 2.21-2.18 (m, 2H), 1.76-1.55 (m, 2H), 1.23-1.20(m, 2H), 0.82 (t, J=7.3 Hz, 3H), 0.61-0.50 (m, 2H), 0.38-0.26 (m, 2H);ESI-MS (m/z): Calculated for: C₂₁H₂₉N₃O₅S: 435.54; observed mass; 434.20(M−H); HPLC purity: 99.1%; R_(t); 7.7

Production Example 76: Synthesis ofN-(1-(3-(cyclopropylmethoxy)phenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

The title compound was prepared using(E)-N-(1-(3-(cyclopropylmethoxy)phenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.082 g, 68%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.18 (s,1H), 7.19 (t, J=7.9 Hz, 1H), 7.00 (t, J=1.9 Hz, 1H), 6.96-6.89 (m, 1H),6.76 (dd, J=8.1, 2.4 Hz, 1H), 3.87 (s, 2H), 3.79 (d, J=7.0 Hz, 2H), 3.12(t, J=7.0 Hz, 2H), 2.54-2.51 (m, 2H), 1.43-1.40 (m, 3H), 1.36-0.95 (m,8H), 0.61-0.50 (m, 2H), 0.35-0.26 (m, 2H); ESI-MS (m/z): Calculated for:C₂₁H₂₉N₃O₅S: 435.54; observed mass: 436.19 (M+H); HPLC purity: 98.0%;R_(t); 7.5

Production Example 78: Synthesis ofN-(1-(3-(2,2-difluoroethoxy)phenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

The title compound was prepared using(E)-N-(1-(3-(2,2-difluoroethoxy)phenyl)cyclopropyl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.082 g, 68%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.18 (s,1H), 7.25 (t, J=7.9 Hz, 1H), 7.07-6.98 (m, 2H), 6.90-6.83 (m, 1H), 6.38(tt, J=56.0, 3.6 Hz, 1H), 4.29 (dt, J=14.7, 3.5 Hz, 2H), 3.87 (s, 2H),3.14 (t, J=9.5, 7.1 Hz, 2H), 2.58-2.54 (m, 2H), 1.45-1.42 (m, 2H),1.30-1.27 (m, 4H), 1.09-1.04 (m, 4H); ESI-MS (m/z): Calculated for:C₁₉H₂₅F₂N₃O₅S: 445.48; observed mass: 446.10 (M+H); HPLC purity: 96.7%;R_(t); 7.3

Exemplary Procedure for the Preparation of2-(3-(cyclopropylmethoxy)-4-fluorophenyl)propan-2-ol (XV)

To a stirred solution of VI (11.0 g, 49.3 mmol) in dry THF (280 mL), wasadded a solution of CH₃MgBr (1.4M soln. in THF, 176.0 mL, 246.6 mmol)drop wise at 0° C. under nitrogen atmosphere and the reaction mixturewas stirred at heated at 80° C. for 3 h. The progress of the reactionwas monitored by TLC. After completion of the reaction, the reactionmixture was quenched with aqueous NH₄Cl and extracted with EtOAc. Thecombined organic layers were washed with brine, dried over anhydrousNa₂SO₄ and evaporated under reduced pressure to afford XV.

Yield: 10.13 g, crude, LCMS: 225.45 (M+1).

Exemplary Procedure for the Preparation of4-(2-azidopropan-2-yl)-2-(cyclopropylmethoxy)-1-fluorobenzene (XVI)

To a stirred solution of XV (10.13 g, 45.8 mmol) in dry DCM (200 mL),NaN₃ (27.00 g 406.8 mmol) and TFA (50 mL) were added at 0° C. and thereaction mixture was stirred at room temperature for 2 h. The progressof the reaction was monitored by TLC. After completion of the reaction,the reaction mixture was quenched with water and extracted with DCM. Thecombined organic layers were washed with brine, dried over anhydrousNa₂SO₄ and evaporated under reduced pressure to afford XVI.

Yield: 10.10 g, crude; ¹H NMR (400 MHz, Chloroform-d) δ 7.10-6.99 (m,2H), 6.96-6.93 (m, 1H), 3.91 (d, J=7.0 Hz, 2H), 1.62-1.56 (m, 6H),1.38-1.19 (m, 2H), 0.71-0.57 (m, 2H), 0.42-0.28 (m, 2H).

Exemplary Procedure for the Preparation of2-(3-(cyclopropylmethoxy)-4-fluorophenyl)propan-2-amine (XVII)

To a stirred solution of XVI (10.0 g, 40.11 mmol) in MeOH (150 mL), 10%Pd/C (4.0 g) was added and stirred under hydrogen atmosphere (balloonpressure) at room temperature for 24 h. The progress of the reaction wasmonitored by TLC. After completion of the reaction, the reaction mixturewas filtered through celite and filtrate was evaporated under reducedpressure. The crude product was purified by column chromatography using5% MeOH in DCM to afford XVII.

Yield: 4.1 g, 45.8%; ¹H NMR (400 MHz, DMSO-d₆) δ 7.28 (dd, J=8.6, 2.2Hz, 1H), 7.11-6.96 (m, 2H), 3.89 (d, J=6.9 Hz, 2H), 1.89-1.83 (m, 2H),1.34 (s, 6H), 0.60-0.58 (m, 2H), 0.34-0.32 (m, 2H).

Exemplary Procedure for the Preparation ofN-(2-(3-(cyclopropylmethoxy)-4-fluorophenyl)propan-2-yl)but-3-ene-1-sulfonamide(XVIII)

To a stirred solution of XVII (1.0 g, 4.47 mmol) in dry DCM (10 mL),Et₃N (1.87 mL, 13.4 mmol) was added and stirred at room temperature for10 min. After that V (1.17 g, 7.61 mmol) in DCM (10 mL) was added dropwise and stirred at room temperature for 2 h. The progress of thereaction was monitored by TLC. After completion of the reaction, thereaction mixture was quenched with water and extracted with EtOAC. Thecombined organic layer were washed with brine, dried over anhydrousNa₂SO₄ and concentrated under reduced pressure. The crude product waspurified by combiflash chromatography using 15% EtOAC/hexane to affordXVIII

Yield: 1.04 g, 64.8%; ¹H NMR (400 MHz, DMSO-d₆) δ 7.48 (s, 1H), 7.26(td, J=7.9, 7.4, 2.3 Hz, 1H), 7.13 (dd, J=11.3, 8.5 Hz, 1H), 7.05-7.0(m, 1H), 5.78-5.63 (m, 1H), 5.04-4.94 (m, 2H), 3.90 (dd, J=7.3, 4.6 Hz,2H), 2.70-2.61 (m, 2H), 2.30 (q, J=7.3 Hz, 2H), 1.59 (d, J=3.2 Hz, 6H),1.24 (td, J=7.8, 4.0 Hz, 1H), 0.63-0.52 (m, 2H), 0.42-0.31 (m, 2H).

Production Example 44: Synthesis of(E)-N-(2-(3-(cyclopropylmethoxy)-4-fluorophenyl)propan-2-yl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

The title compound was prepared usingN-(2-(3-(cyclopropylmethoxy)-4-fluorophenyl)propan-2-yl)but-3-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.095 g, 20.65%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (s, 1H), 7.47(s, 1H), 7.26 (dd, J=8.3, 2.3 Hz, 1H), 7.12 (dd, J=11.2, 8.5 Hz, 1H),7.06-6.97 (m, 1H), 5.56-5.53 (m, 1H), 5.39-5.36 (m, 1H), 3.92-3.80 (m,2H), 3.76 (s, 2H), 3.70-3.68 (m, 2H), 2.74-2.65 (m, 2H), 2.33-2.24 (m,2H), 1.58 (s, 6H), 1.24-1.20 (m, 1H), 0.63-0.53 (m, 2H), 0.37-0.29 (m,2H); ESI-MS (m/z): Calculated for: C₂₁H₂₈FN₃O₅S: 453.53: observed mass;471.25 (M+H₂O); HPLC purity: 99.8%; R_(t); 8.0

Production Example 48: Synthesis of(E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(2-(4-fluoro-3-isobutoxyphenyl)propan-2-yl)pent-3-ene-1-sulfonamide

The title compound was prepared usingN-(2-(4-fluoro-3-isobutoxyphenyl)propan-2-yl)but-3-ene-1-sulfonamide and1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.056 g, 11%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (s, 1H), 7.49 (d,J=4.7 Hz, 1H), 7.29 (d, J=8.4 Hz, 1H), 7.12 (dd, J=11.3, 8.1 Hz, 1H),7.01 (dd, J=7.7, 4.0 Hz, 1H), 5.54-5.51 (m, 1H), 5.37-5.34 (m, 1H),3.89-3.73 (m, 6H), 2.70-2.67 (m, 2H), 2.30-2.26 (m, 2H), 2.08-2.04 (m,1H), 1.59 (s, 6H), 0.99 (d, J=6.6 Hz, 6H); ESI-MS (m/z): Calculated for:C₂₁H₃₀F₃O₅S: 455.55; observed mass; 454.20 (M−H); HPLC purity: 98.9%;R_(t); 8.2

Production Example 81: Synthesis ofN-(2-(3-(cyclopropylmethoxy)-4-fluorophenyl)propan-2-yl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

The title compound was prepared using(E)-N-(2-(3-(cyclopropylmethoxy)-4-fluorophenyl)propan-2-yl)-5-(2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.05 g, 71.4%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 7.41 (s,1H), 7.26 (dd, J=8.3, 2.3 Hz, 1H), 7.13 (dd, J=11.3, 8.5 Hz, 1H),7.05-7.01 (m, 1H), 3.89-3.85 (m, 4H), 3.21-3.13 (m, 2H), 2.66-2.57 (m,2H), 1.56-1.54 (m, 8H), 1.40-1.38 (m, 2H), 1.31-1.12 (m, 3H), 0.63-0.53(m, 2H), 0.38-0.29 (m, 2H); ESI-MS (m/z): Calculated for: C₂₁H₃₀FN₃O₅S:455.55; observed mass; 473.15 (M+H₂O); HPLC purity: 94.7%; R_(t); 7.9

Production Example 86: Synthesis of5-(2,4-dioxoimidazolidin-1-yl)-N-(2-(4-fluoro-3-isobutoxyphenyl)propan-2-yl)pentane-1-sulfonamide

The title compound was prepared using(E)-5-(2,4-dioxoimidazolidin-1-yl)-N-(2-(4-fluoro-3-isobutoxyphenyl)propan-2-yl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.028 g, 70%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 7.42 (s,1H), 7.32-7.24 (m, 1H), 7.12 (dd, J=11.4, 8.4 Hz, 1H), 7.00-6.98 (m,1H), 3.89 (s, 2H), 3.81 (d, J=6.5 Hz, 2H), 3.17 (t, J=7.1 Hz, 2H),2.50-2.48 (m, 2H), 2.05-2.01 (m, 1H), 1.56-1.54 (m, 8H), 1.40-1.37 (m,2H), 1.20-1.00 (m, 2H), 0.99 (d, J=6.6 Hz, 6H); ESI-MS (m/z): Calculatedfor: C₂₁H₃₂FN₃O₅S: 457.56; observed mass: 479.85 (M+Na); HPLC purity:96.3%; R_(t); 8.2

Exemplary Procedure for the Preparation of N-allylcyanamide (XIX)

To a stirred solution of methyl L-alaninate (5.0 g, 35 mmol) in dry ACN(25 mL), Et₃N (10.0 mL, 71.0 mmol) was added and stirred at roomtemperature for 10 min. After that 3-bromoprop-1-ene (2.78 mL, 32.0mmol) in ACN (25 mL) was added at 0° C. drop wise and stirred at roomtemperature for 16 h. The progress of the reaction was monitored by TLC.After completion of the reaction, the reaction mixture was quenched withwater and extracted with DCM. The combined organic layer were washedwith brine, dried over anhydrous Na₂SO₄ and concentrated under reducedpressure. The crude product was purified by column chromatography using10% MeOH/DCM to afford XIX.

Yield: 0.8 g, 15.6%; ¹H NMR (400 MHz, DMSO-d₆) δ 5.80-5.77 (m, 1H),5.22-4.98 (m, 2H), 4.03 (d, J=7.1 Hz, 1H), 3.35-3.10 (m, 3H), 3.10-2.98(m, 1H), 2.13-2.08 (m, 1H), 1.99 (s, 1H), 1.17 (dd, J=7.1, 5.3 Hz, 5H).

Exemplary Procedure for the Preparation of methylN-allyl-N-cyano-L-alaninate (XX)

To a stirred solution of XIX (0.8 g, 5.5 mmol) in EtO₂ (10 mL), BrCN(7.11 g, 6.7 mmol) and NaHCO₃ (1.40 g, 16.6 mmol) were added dropwise at0° C. and stirred for 2 h. The progress of the reaction was monitored byTLC. After completion of the reaction, the reaction mixture was quenchedwith water and extracted with EtO₂. The combined organic layer werewashed with brine, dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to afford XX.

Yield: 0.8 g, crude; NMR: ¹H NMR (400 MHz, DMSO-d₆) δ 5.84-5.81 (m, 1H),5.36-5.21 (m, 2H), 3.93 (d, J=7.3 Hz, 1H), 3.79-3.58 (m, 5H), 1.46-1.34(m, 3H).

Exemplary Procedure for the Preparation of(S)-1-allyl-5-methylimidazolidine-2,4-dione (XXI)

To a stirred solution of XX (0.8 g, 4.7 mmol) in toluene (8 mL), dibutylphosphate (2.5 mL, 1.1 mmol) was added and reaction mixture was heatedto reflux for 5 h. The progress of the reaction was monitored by TLC.After completion of the reaction, reaction mixture was concentrated andthe residue was dissolved in Et₂O: hexane (2:8 mL), precipitated solidwas filtered and which was purified by trituration with cold hexane toafford XXI.

Yield: 0.25 g, 34%; NMR: ¹H NMR (400 MHz, DMSO-d₆) δ 10.80 (s, 1H),5.80-5.77 (m, 1H), 5.27-5.11 (m, 2H), 4.14-3.94 (m, 2H), 3.70 (dd,J=16.1, 6.3 Hz, 1H), 1.41-1.22 (m, 3H).

Production Example 15: Synthesis of (R,E)-N-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)-5-(5-methyl-2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

The title compound was prepared usingN-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)but-3-ene-1-sulfonamide and(R)-1-allyl-5-methylimidazolidine-2,4-dione in a manner similar to themethod in Production Example 3 above.

Yield: 0.028 g, 18%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (s, 1H), 8.23 (s,1H), 7.23 (d, J=8.0 Hz, 1H), 7.15-7.05 (m, 1H), 6.92 (s, 1H), 5.48-5.36(m, 1H), 5.24-5.20 (m, 1H), 3.98-3.86 (m, 2H), 3.83-3.76 (m, 2H),3.51-3.48 (m, 1H), 2.57-2.54 (m, 2H), 2.19-1.95 (m, 4H), 1.29-1.12 (m,4H), 1.07 (s, 2H), 1.01-0.94 (m, 6H); ESI-MS (m/z): Calculated for:C₂₂H₃₀FN₃O₅S: 467.56; observed mass; 468.15 (M+H); HPLC purity: 94.7%;R_(t); 8.2

Production Example 16: Synthesis of (S,E)-N-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)-5-(5-methyl-2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

The title compound was prepared usingN-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)but-3-ene-1-sulfonamide and(S)-1-allyl-5-methylimidazolidine-2,4-dione in a manner similar to themethod in Production Example 3 above.

Yield: 0.025 g, 18%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (s, 1H), 8.23 (s,1H), 7.28-7.06 (m, 2H), 6.92 (d, J=7.6 Hz, 1H), 5.49-5.37 (m, 1H),5.26-5.23 (m, 1H), 4.05-3.87 (m, 2H), 3.81 (d, J=6.4 Hz, 2H), 3.53-3.50(m, 1H), 2.59-2.50 (m, 3H), 2.10-2.07 (m, 2H), 1.27-1.16 (m, 5H),1.09-1.04 (m, 2H), 0.99 (d, J=6.6 Hz, 6H); ESI-MS (m/z): Calculated for:C₂₂H₃₀FN₃O₅S: 467.56; observed mass: 468.20 (M+H); HPLC purity: 99.8%;R_(t); 8.2

Production Example 17: Synthesis of (R,E)-N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)cyclopropyl)-5-(5-methyl-2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

The title compound was prepared usingN-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)cyclopropyl)but-3-ene-1-sulfonamideand (R)-1-allyl-5-methylimidazolidine-2,4-dione in a manner similar tothe method in Production Example 3 above.

Yield: 0.038 g, 14%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (s, 1H), 8.22 (s,1H), 7.20 (dd, J=8.3, 2.2 Hz, 1H), 7.11 (dd, J=11.4, 8.4 Hz, 1H),6.96-6.91 (m, 1H), 5.49-5.37 (m, 1H), 5.30-5.18 (m, 1H), 3.99-3.85 (m,4H), 3.53 (dd, J=15.6, 6.9 Hz, 1H), 2.59-2.56 (m, 2H), 2.14-2.11 (m,2H), 1.29-1.17 (m, 6H), 1.11-1.03 (m, 2H), 0.63-0.53 (m, 2H), 0.38-0.29(m, 2H); ESI-MS (m/z): Calculated for: C₂₂H₂₈FN₃O₅S: 465.54; observedmass; 466.15 (M+H); HPLC purity: 99.9%; R_(t); 7.8

Production Example 18: Synthesis of (R,E)-N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)cyclopropyl)-5-(5-methyl-2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamide

The title compound was prepared usingN-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)cyclopropyl)but-3-ene-1-sulfonamideand (S)-1-allyl-5-methylimidazolidine-2,4-dione in a manner similar tothe method in Production Example 3 above.

Yield: 0.06 g, 23%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (s, 1H), 8.12 (s,1H), 7.26 (dt, J=7.9, 7.4, 2.3 Hz, 1H), 7.13 (dd, J=11.3, 8.5 Hz, 1H),7.05-7.0 (m, 1H), 5.45-5.38 (m, 1H), 5.12-5.08 (m, 1H), 3.99-3.85 (m,4H), 3.37-3.25 (m, 1H), 2.59 (t, J=7.8 Hz, 2H), 2.15 (t, J=7.7 Hz, 2H),1.26-1.17 (m, 6H), 1.08-1.04 (m, 2H), 0.63-0.53 (m, 2H), 0.36-0.30 (m,2H); ESI-MS (m/z): Calculated for C₂₂H₂₈FN₃O₅S: 465.54; observed mass;464.20 (M−H); HPLC purity: 99.4%; R_(t); 7.7

Production Example 71: Synthesis of(R)—N-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)-5-(5-methyl-2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

The title compound was prepared usingN-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)but-3-ene-1-sulfonamide ina manner similar to the method in Production Example 1 above.

Yield: 0.022 g, 62%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (d, J=11.1 Hz,1H), 8.19 (d, J=13.7 Hz, 1H), 7.25 (dd, J=8.5, 2.2 Hz, 1H), 7.12 (dd,J=11.4, 8.3 Hz, 1H), 6.96-6.90 (m, 1H), 4.01-3.38 (m, 1H), 3.81 (d,J=6.6 Hz, 2H), 2.96-2.91 (m, 1H), 2.57-2.54 (m, 3H), 2.06-2.02 (m, 1H),1.49-1.18 (m, 8H), 1.11-0.92 (m, 11H); ESI-MS (m/z): Calculated for:C₂₂H₃₂FN₃O₅S: 469.57; observed mass; 470.20 (M+H); HPLC purity: 92.2%;Rt; 8.5

Production Example 72: Synthesis of(S)—N-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)-5-(5-methyl-2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

The title compound was prepared usingN-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)but-3-ene-1-sulfonamide ina manner similar to the method in Production Example 1 above.

Yield: 0.042 g, 83.6%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.18(s, 1H), 7.25 (dd, J=8.4, 2.3 Hz, 1H), 7.12 (dd, J=11.4, 8.4 Hz, 2H),6.94-6.90 (m, 1H), 4.01-3.39 (m, 1H), 3.81 (d, J=6.6 Hz, 2H), 3.32-3.28(m, 2H), 2.98-2.95 (m, 2H), 2.06-2.03 (m, 2H), 1.43-1.40 (m, 2H),1.36-1.18 (m, 8H), 1.11-0.95 (m, 7H); ESI-MS (m/z): Calculated for:C₂₂H₃₂FN₃O₅S: 469.57; observed mass; 470.2 (M+H); HPLC purity: 98.0%;R_(t): 8.3

Production Example 73: Synthesis of(R)—N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)cyclopropyl)-5-(5-methyl-2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

The title compound was prepared usingN-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)but-3-ene-1-sulfonamide ina manner similar to the method in Production Example 1 above.

Yield: 0.02 g, 80.0%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.15 (s,1H), 7.20 (d, J=8.2 Hz, 1H), 7.11 (t, J=10.1 Hz, 1H), 6.96-6.92 (m, 1H),4.03-4.00 (m, 1H), 3.88 (d, J=7.0 Hz, 2H), 2.98-2.92 (m, 1H), 2.51-2.40(m, 2H), 1.41-1.38 (m, 2H), 1.28-1.17 (m, 9H), 1.04-1.00 (m, 4H),0.60-0.55 (m, 2H), 0.32-0.30 (m, 2H); ESI-MS (m/z): Calculated for:C₂₂H₃₀F₃O₅S: 467.56; observed mass; 468.20 (M+H); HPLC purity: 94.1%;R_(t): 7.7

Production Example 74: Synthesis of(S)—N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)cyclopropyl)-5-(5-methyl-2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

The title compound was prepared using (S,E)-N-(1-(4-fluoro-3-isobutoxyphenyl)cyclopropyl)-5-(5-methyl-2,4-dioxoimidazolidin-1-yl)pent-3-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.035 g, 77.7%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 8.16(s, 1H), 7.21 (d, J=8.2 Hz, 1H), 7.12 (dd, J=11.4, 8.2 Hz, 1H),6.97-6.89 (m, 1H), 4.02-3.98 (m, 1H), 3.88 (d, J=6.9 Hz, 2H), 2.99-2.87(m, 2H), 2.58-2.48 (m, 2H), 1.48-1.18 (m, 12H), 1.05-1.02 (m, 2H),0.59-0.54 (m, 2H), 0.33-0.31 (m, 2H); ESI-MS (m/z): Calculated for:C₂₂H₃₀F₃O₅S: 467.56; observed mass; 468.15 (M+H); HPLC purity: 96.9%;R_(t): 7.7

Exemplary Procedure for the preparation of3-(cyclopropylmethoxy)-4-fluoro-N-methoxy-N-methylbenzamide (XXII)

To a mixture of 3-(cyclopropylmethoxy)-4-fluorobenzoic acid (10.12 g,4.80 mmol) in dry DMF (131 mL), N,O-dimethylhydroxylamine (5.63 g, 5.70mmol), HOBt (7.69 g, 5.70 mmol), Et₃N (8.75 ml, 6.20 mmol) and EDCI.HCl(13.85 g, 7.20 mmol) were added at 0° C. and reaction mixture wasstirred at room temperature for 16 h. The progress of the reaction wasmonitored by TLC. After completion of the reaction, the reaction mixturewas quenched with water and extracted with EtOAc. The combined organiclayer were washed with brine, dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The residue was purified by columnchromatography using 40% EtOAc/hexane to afford XXII

Yield: 10.5 g, 86.1%; ¹H NMR (400 MHz, CDCl₃) δ 7.39-7.24 (m, 2H), 7.08(dd, J=10.9, 8.4 Hz, 1H), 3.90 (d, J=7.0 Hz, 2H), 3.55 (s, 3H), 3.35 (s,3H), 1.37-1.23 (m, 1H), 0.72-0.59 (m, 2H), 0.43-0.29 (m, 2H).

Exemplary Procedure for the Preparation of1-(3-(cyclopropylmethoxy)-4-fluorophenyl)propan-1-one (XXIII)

To a stirred solution of XXII (10.5 g, 41.40 mmol) in dry THF (158 mL),was added a solution of CH₃CH₂MgBr (1.0M soln. in THF, 34.5 mL, 103.0mmol) drop wise at 0° C. and the reaction mixture was stirred at roomtemperature for 3 h. The progress of the reaction was monitored by TLC.After completion of the reaction, the reaction mixture was quenched withNH₄Cl solution and extracted with EtOAc. The combined organic layerswere washed with brine, dried over anhydrous Na₂SO₄ and evaporated underreduced pressure. The residue was purified by column chromatographyusing 25% EtOAc/hexane to afford XXIII

Yield: 7.58 g, 82.3%; ESI-MS (m/z): 222.85 (M+H).

Exemplary Procedure for the Preparation of4-(but-1-en-2-yl)-2-(cyclopropylmethoxy)-1-fluorobenzene (XXIV)

To a stirred solution of Ph₃PCH₃Br (17.93 g, 50.2 mmol) in dry THF (120mL), NaHMDS (1M in THF 50 mL, 50.2 mmol) was added 0° C. and stirred atroom temperature for 2 h. XXIII (6.2 g, 27.8 mmol) in THF (50 mL) wasadded drop wise 0° C. and reaction was stirred at room temperature for12 h. The progress of the reaction was monitored by TLC. Aftercompletion of the reaction, the reaction mixture was quenched with NH₄Clsolution and extracted with EtOAc. The combined organic layer werewashed with brine, dried over anhydrous Na₂SO₄ and concentrated underreduced pressure. The residue was purified by column chromatographyusing 5% EtOAc/hexane to afford XXIV.

Yield: 5.6 g, 91.2%; ¹H NMR (400 MHz, CDCl₃) δ 7.06-6.88 (m, 3H), 5.19(s, 1H), 5.05-5.00 (m, 1H), 3.89 (d, J=7.0 Hz, 2H), 2.46 (q, J=7.3 Hz,2H), 1.33-1.30 (m, 1H), 1.09 (td, J=7.4, 1.3 Hz, 3H), 0.70-0.58 (m, 2H),0.43-0.32 (m, 2H).

Exemplary Procedure for the Preparation of(S)-2-(3-(cyclopropylmethoxy)-4-fluorophenyl)butane-1,2-diol (XXV)

To a mixture of XXIV (3.0 g, 13.6 mmol) in t-Butanol (48 mL) and water(48 mL), AD-mix-alpha (18.0 g) was added at 0° C. and reaction mixturewas stirred at room temperature for 3 h. The progress of the reactionwas monitored by TLC. After completion of the reaction, the reactionmixture was quenched with anhydrous Na₂SO₄ and extracted with EtOAc. Thecombined organic layer were washed with brine, dried over anhydrousNa₂SO₄ and concentrated under reduced pressure. The residue was purifiedby column chromatography using 35% EtOAc/hexane to afford XXV.

Yield: 2.8 g, 80.92%; ¹H NMR (400 MHz, CDCl₃) δ 7.13-7.00 (m, 2H),6.88-6.85 (m, 1H), 3.90 (d, J=7.0 Hz, 2H), 3.81 (dd, J=11.1, 4.6 Hz,1H), 3.67 (dd, J=11.0, 8.0 Hz, 1H), 2.54 (s, 1H), 1.83-1.79 (m, 2H),1.58 (dd, J=8.1, 4.7 Hz, 1H), 1.29-1.27 (m, 2H), 0.77 (dd, J=8.0, 6.8Hz, 3H), 0.70-0.58 (m, 2H), 0.37 (t, J=5.2 Hz, 2H).

Exemplary Procedure for the Preparation of(S)-2-(3-(cyclopropylmethoxy)-4-fluorophenyl)-1-(methylsulfonyl)butan-2-ol(XXVI)

To a stirred solution of XXV (0.86 g, 3.38 mmol) and Et₃N (0.711 mL,5.07 mmol) in dry DCM (8.6 mL), MsCl (0.31 mL, 4.06 mmol) was added at0° C. and stirred at room temperature for 30 min. After completion ofthe reaction, the reaction mixture was quenched with NaHCO₃ solution andextracted with EtOAc. The combined organic layer were washed with brine,dried over anhydrous Na₂SO₄ and concentrated under reduced pressure toafford XXVI.

Yield: 1.0 g, crude.

Exemplary Procedure for the Preparation of(S)-1-azido-2-(3-(cyclopropylmethoxy)-4-fluorophenyl)butan-2-ol (XXVII)

To a stirred solution of XXVI (0.75 g, 2.20 mmol) in DMF (15 mL), sodiumazide (0.586, 9.02 mmol) was added drop wise and stirred at 90° C. for12 h. The progress of the reaction was monitored by TLC. Aftercompletion of the reaction, the reaction mixture was diluted with waterand extracted with EtOAc. The combined organic layers were washed withbrine, dried over anhydrous Na₂SO₄ and concentrated under reducedpressure. The residue was purified by column chromatography using 35%EtOAc/hexane to afford XXVII.

Exemplary Procedure for the Preparation of(S)-1-amino-2-(3-(cyclopropylmethoxy)-4-fluorophenyl)butan-2-ol (XXVIII)

To a stirred solution of XXVII (0.3 g, 10.7 mmol) in EtOH (6 mL), 10%Pd/C (0.06 g) was added and stirred under hydrogen atmosphere (balloonpressure) at room temperature for 3 h. The progress of the reaction wasmonitored by TLC. After completion of the reaction, the reaction mixturewas filtered through celite and filtrate was evaporated under reducedpressure. The residue was purified by column chromatography using 40%EtOAc/hexane to afford XXVIII.

Yield: 0.097 g, 34.7%; ¹H NMR (400 MHz, DMSO-d₆) δ 7.15-7.05 (m, 2H),6.93-6.89 (m, 1H), 4.82 (s, 1H), 3.87 (d, J=7.1 Hz, 2H), 2.79 (d, J=13.1Hz, 1H), 2.70 (d, J=13.1 Hz, 1H), 1.75-1.61 (m, 2H), 1.25-1.22 (m, 1H),0.68-0.50 (m, 5H), 0.40-0.26 (m, 2H).

Exemplary Procedure for the Preparation of(S)—N-(2-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-hydroxybutyl)prop-2-ene-1-sulfonamide(XXIX)

To a stirred solution of XXVIII (1. g, 7.1 mmol) in dry DCM (25 mL),Et₃N (3.0 mL, 2.1 mmol) was added and stirred at room temperature for 10min. After that V (1.49 g, 1.0 mmol) in DCM (25 mL) was added drop wiseand stirred at room temperature for 12 h. The progress of the reactionwas monitored by TLC. After completion of the reaction, the reactionmixture was quenched with water and extracted with EtOAc. The combinedorganic layer were washed with brine, dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The residue was purified by columnchromatography using 20-30% EtOAc/hexane to afford XXIX.

Yield: 0.8 g, 32%; ¹H NMR (400 MHz, DMSO-d₆) δ 7.16-7.11 (m, 2H),6.95-6.91 (m, 1H), 6.70 (t, J=6.0 Hz, 1H), 5.72-5.69 (m, 1H), 5.34-5.25(m, 2H), 4.94 (d, J=2.0 Hz, 1H), 3.92-3.85 (m, 2H), 3.65-3.60 (m, 2H),3.39-3.15 (m, 1H), 1.74 (q, J=7.5 Hz, 2H), 1.25-1.21 (m, 2H), 0.66-0.52(m, 5H), 0.33 (t, J=4.3 Hz, 2H).

Production Example 97: Synthesis of (S,E)-N-(2-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-hydroxybutyl)-4-(2,4-dioxoimidazolidin-1-yl)but-2-ene-1-sulfonamide

The title compound was prepared using(S)—N-(2-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-hydroxybutyl)prop-2-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.037 g, 7.1%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (s, 1H),7.13-7.10 (m, 2H), 6.96-6.88 (m, 1H), 6.79-6.76 (m, 1H), 5.71-5.68 (m,1H), 5.53-5.50 (m, 1H), 4.94 (s, 1H), 3.88-3.84 (m, 6H), 3.70-3.68 (m,2H), 3.25-3.20 (m, 2H), 1.73-1.70 (m, 2H), 1.22-1.20 (m, 1H), 0.65-0.54(m, 5H), 0.33-0.30 (m, 2H); ESI-MS (m/z): Calculated for: C₂₁H₂₈FN₃O₆S:469.53; observed mass; 492.20 (M+Na); HPLC purity: 96.5%; R_(t): 7.7

Production Example 82: Synthesis of (S,E)-4-(2,4-dioxoimidazolidin-1-yl)-N-(2-(4-fluoro-3-isobutoxyphenyl)-2-hydroxybutyl)but-2-ene-1-sulfonamide

The title compound was prepared using (S,E)-4-(2,4-dioxoimidazolidin-1-yl)-N-(2-(4-fluoro-3-isobutoxyphenyl)-2-hydroxybutyl)but-2-ene-1-sulfonamideand 1-allylimidazolidine-2,4-dione in a manner similar to the method inProduction Example 3 above.

Yield: 0.022 g, 3.67%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.8 (s, 1H),7.18-7.06 (m, 2H), 6.98-6.92 (m, 1H), 6.77 (t, J=6.3 Hz, 1H), 5.70-5.67(m, 1H), 5.52-5.49 (m, 1H), 4.96 (s, 1H), 3.87-3.77 (m, 6H), 3.70-3.68(m, 2H), 3.30-3.28 (m, 2H), 2.11-1.96 (m, 1H), 1.74-1.71 (m, 2H), 0.98(d, J=6.6 Hz, 6H), 0.60 (t, J=7.4 Hz, 3H); ESI-MS (m/z): Calculated for:C₂₁H₃₀F₃O₆S: 471.54; observed mass; 489.30 (M+H₂O); HPLC purity: 94.7%;R_(t): 7.9

Production Example 98: Synthesis of(S)—N-(2-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-hydroxybutyl)-4-(2,4-dioxoimidazolidin-1-yl)butane-1-sulfonamide

The title compound was prepared using (S,E)-N-(2-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-hydroxybutyl)-4-(2,4-dioxoimidazolidin-1-yl)but-2-ene-1-sulfonamidein a manner similar to the method in Production Example 1 above.

Yield: 0.06 g, 48%; ¹H NMR (400 MHz, DMSO-d₆) δ 10.7 (s, 1H), 7.15-7.11(m 2H), 6.97-6.89 (m, 1H), 6.65 (t, J=6.2 Hz, 1H), 4.92 (s, 1H),3.89-3.86 (m, 4H), 3.20-3.17 (m, 4H), 2.89-2.86 (m, 2H), 1.74-1.71 (m,2H), 1.48-1.41 (m, 4H), 1.29-1.13 (m, 1H), 0.60-0.56 (m, 5H), 0.33-0.30(m, 2H); ESI-MS (m/z): Calculated for: C₂₁H₃₀FN₃O₆S; observed mass;470.25 (M−H); HPLC purity: 97.9; R_(t): 7.3.

Production Example 66a: Synthesis of(S)—N-(1-(4-chloro-3-(cyclopropylmethoxy) phenyl) ethyl)-5-(2,4-dioxoimidazolidin-1-yl) pentane-1-sulfonamide

Step-1: Synthesis of 1-(4-chloro-3-(cyclopropylmethoxy) phenyl)ethan-1-one (3-a)

To a stirred solution of 1-a (3.0 g, 17.6 mmol) in DMF (25 mL) was addedK₂CO₃ (7.3 g, 52.9 mmol) followed by addition of(bromomethyl)cyclopropane (2.8 g, 21.1 mmol) and the reaction mixturewas refluxed for 4 h (reaction deemed complete by TLC). The reactionmixture was quenched with water and extracted with EtOAc. The combinedorganic layer was washed with brine, dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure to afford 3-a (3.2 g, crude).

Step-2: Synthesis ofN-(1-(4-chloro-3-(cyclopropylmethoxy)phenyl)ethylidene)-2-methylpropane-2-sulfinamide(5-a)

To a stirred solution of compound 3-a (3.2 g, 14.2 mmol) and compound4-a (2.5 g, 21.4 mmol) in dry toluene (150 mL) was added Ti(O^(i)Pr)₄(8.1 g, 28.5 mmol). The resultant mixture was heated at 90° C. for 16 h(reaction deemed complete by TLC). The reaction mixture was diluted withEtOAc and quenched with water and filtered through a pad of Celite. Thefiltrate was diluted with water and extracted with EtOAc. The combinedorganic layer was washed with brine, dried over anhydrous Na₂SO₄ andevaporated under reduced pressure. The residue was purified by columnchromatography using 10% EtOAc/hexane to afford compound 5-a (2.4 g,crude). LCMS: 327.95 (M+1).

Step-3: Synthesis of (S)—N—((S)-1-(4-chloro-3-(cyclopropylmethoxy)phenyl) ethyl)-2-methylpropane-2-sulfinamide (6-a)

To a stirred solution of DIBAL-H (1M solution in toluene, 22 mL, 22.0mmol) in dry toluene (10 mL), was added a solution of compound 5-a (2.4g, 7.33 mmol) in toluene (10 mL) dropwise at −78° C. The resultantmixture was stirred at same temperature for 3 h (reaction deemedcomplete by TLC). The reaction mixture was quenched with NH₄Cl solutionand diluted with EtOAc. The reaction mixture was filtered through a padof Celite and extracted with EtOAc, washed with brine, dried overanhydrous Na₂SO₄ and evaporated under reduced pressure. The residue waspurified by column chromatography using 10% EtOAc/hexane to affordcompound 6-a (1.6 g, 66%). LCMS: 330.1 (M+1).

Step-4: Synthesis of (S)-1-(4-fluoro-3-propoxyphenyl) ethan-1-aminehydrochloride (7-a)

To a stirred solution of compound 6-a (1.6 g, 4.86 mmol) in MeOH (15mL), was added 4M HCl in dioxane (2.4 mL, 9.72 mmol) and the resultingmixture stirred at room temperature for 3 h (reaction deemed complete byTLC). The reaction mixture was concentrated and the residue was purifiedby trituration with diethyl ether to afford 7-a (0.9 g, 75%). LCMS:208.9 (M-18).

¹H NMR (400 MHz, DMSO-d₆) δ 8.42 (s, 3H), 7.42 (d, J=8.7 Hz 1H), 7.20(s, 1H), 4.42-4.38 (m, 1H), 3.96 (t, J=7.0 Hz, 2H), 1.45-1.42 (s, 3H),1.28-1.0 (m, 2H), 0.62-0.56 (m, 2H), 0.38-0.34 (m, 2H).

Step 5: Synthesis of3-((2-(trimethylsilyl)ethoxy)methyl)imidazolidine-2,4-dione (9-a)

To a stirred solution of compound 8-a (6.0 g, 60.0 mmol) and DIPEA (30mL, 180 mmol) in CH₂Cl₂ (60 mL) was added SEM-Cl (12.7 mL, 72.0 mmol) at0° C. over a period of 1 h and stirred at the room temperature for 16 h.The reaction was monitored by TLC for complete consumption of compound8-a. The reaction mixture was quenched with NH₄Cl solution (150 mL) andextracted with CH₂Cl₂ (150 mL×2). The combined organic extracts waswashed with 2N HCl (75 mL×2) and brine (100 mL), dried over anhydroussodium sulfate and concentrated under reduced pressure to affordcompound 9-a (9.2 g, crude).

¹H NMR (400 MHz, DMSO-d₆) δ 4.98 (s, 2H), 4.0 (s, 3H), 3.60-3.55 (m,2H), 0.98-0.95 (m, 2H), 0.09 (s, 9H).

Step 6: Synthesis of (1-(5-bromopentyl)-3-((2-(trimethylsilyl) ethoxy)methyl) imidazolidine-2, 4-dione (11-a)

To a stirred solution of compound 9-a (3.0 g, 13.0 mmol) in ACN (50 mL)was added CS₂CO₃ (12.7 g, 39.1 mmol) followed by addition of compound10-a (6.0 g, 26.0 mmol) and stirred at 80° C. for 4 h. After completion,Reaction mixture was concentrated under reduced pressure and residue wastaken in saturated solution of water (150 mL) and extracted with EtOAc(150 mL×2). The organic extract was washed with brine (150 mL)respectively then dried over anhydrous sodium sulfate and concentratedunder reduced pressure to dryness. The residue was purified bycombiflash chromatography (eluting with 3-4% MeOH in DCM) to afford 11-a(2.8 g, 56%).

¹H NMR (400 MHz, DMSO-d₆) δ 4.73 (s, 2H), 4.02 (s, 2H), 3.52-3.50 (m,4H), 3.22-3.20 (m, 2H), 1.85-1.79 (m, 2H), 1.59-1.55 (m, 2H), 1.41-1.34(m, 2H), 0.88-0.82 (m, 2H), 0.09 (s, 9H).

Step 7: Synthesis ofS-(5-(2,4-dioxo-3-((2-(trimethylsilyl)ethoxy)methyl)imidazolidin-1-yl)pentyl)ethanethioate (12-a)

To a solution of compound 11-a (2.8 g, 7.38 mmol) in DMF (30 mL) wasadded AcSK (1 g, 8.86 mmol) at 0° C. and stirred at room temperature for1 h. After completion of starting material, the reaction mixture wasdiluted with water (75 mL) and extracted with EtOAc (75 mL×3). Theorganic extract was washed with brine (50 mL) respectively then driedover anhydrous sodium sulfate and concentrated under reduced pressure toafford 12-a (2.6 g, crude). It was carried forward to the next stepwithout purification.

Step 8: Synthesis of 5-(2, 4-dioxo-3-((2-(trimethylsilyl) ethoxy)methyl) imidazolidin-1-yl) pentane-1-sulfonyl chloride (13-a)

2N HCl (4 mL) was added to a stirred solution of compound 12-a (2.6 g,6.87 mmol) in acetonitrile (40 mL) at 0° C. This was followed byportion-wise addition of N-chlorosuccinimide (3.67 g, 27.5 mol) over 30min and the reaction mixture was warmed to room temperature, stirred for1 h. Reaction progress was monitored by TLC. Upon complete consumptionof 12-a, reaction mixture was quenched with ice-cold water (150 mL) andextracted with diethyl ether (150 mL×2). The combined organic extractswas washed with saturated sodium bicarbonate solution (150 mL) and brine(75 mL) and dried over anhydrous sodium sulfate and concentrated underreduced pressure to afford 13-a as an off-white solid (1.9 g, 69.3%).

¹H NMR (400 MHz, DMSO) δ 4.90 (s, 2H), 3.89 (s, 2H), 3.69-3.59 (m, 2H),3.46-3.42 (m, 2H), 2.13-2.0 (m, 2H), 1.70-1.64 (m, 2H), 1.62-1.52 (m,2H), 1.27-1.23 (m, 2H), 0.96-0.92 (m, 2H), 0.004 (s, 9H).

Step 9: Synthesis of(S)—N-(1-(4-chloro-3-(cyclopropylmethoxy)phenyl)ethyl)-5-(2,4-dioxo-3-((2-(trimethylsilyl)ethoxy)methyl)imidazolidin-1-yl)pentane-1-sulfonamide(14-a)

To a stirred solution of compound 7-a —HCl salt (0.2 g, 0.76 mmol) inCH₂Cl₂ (5 mL) was added triethylamine (0.25 mL, 2.28 mmol) at 0° C. andstirred. To this reaction mixture was added compound 13-a (0.33 g, 0.83mmol) in CH₂Cl₂ (5 mL) dropwise over 25 min at 0° C. and stirred at thesame temperature for 3 h. The reaction was monitored by TLC for completeconsumption of compound 7-a. The reaction mixture was quenched withwater (25 mL) and extracted with CH₂Cl₂ (25 mL×2). The combined organicextracts was washed with water (25 mL×2) and brine (25 mL), dried overanhydrous sodium sulfate and concentrated under reduced pressure. Thecrude was purified by silica gel flash chromatography using 15-20%EtOAc/hexanes to afford 14-a as an off-white solid (0.155 g, 34%). LCMS:588.05 (M+1).

Step 10: Synthesis of (S)—N-(1-(4-chloro-3-(cyclopropylmethoxy) phenyl)ethyl)-5-(2, 4-dioxoimidazolidin-1-yl) pentane-1-sulfonamide (ProductionExample 66a)

To a stirred solution of compound 14-a (0.15 g, 0.52 mmol) in DCM (10mL) was added TFA (0.3 mL) at 0° C. and the reaction mixture was stirredat room temperature for 16 h. The progress of the reaction was monitoredby TLC till complete consumption of compound 14-a. Reaction mixture wasconcentrated under reduced pressure and residue was taken in saturatedsolution of NaHCO₃ solution (50 mL) and extracted with EtOAc (50 mL×2).The organic extract was washed with water (50 mL×2) and brine (10 mL)respectively then dried over anhydrous sodium sulfate and concentratedunder reduced pressure to dryness. The residue was purified bycombiflash chromatography (eluting with 3-4% MeOH in DCM) to affordProduction Example 66a as a white sticky solid (75 mg, 64% yield).

¹H NMR (400 MHz, DMSO-d₆) δ 10.68 (s, 1H), 7.69 (d, J=8.7 Hz, 1H), 7.35(d, J=8.0 Hz, 1H), 7.16 (s, 1H), 6.93 (d, J=8.1 Hz, 1H), 4.40-4.35 (m,1H), 3.94-3.84 (m, 4H), 3.12 (t, J=7.0 Hz, 2H), 2.82-2.77 (m, 1H),2.63-2.60 (m, 1H), 1.59-1.38 (m, 2H), 1.38-0.99 (m, 8H), 0.58 (d, J=7.8Hz, 2H), 0.35 (d, J=7.6 Hz, 2H); ESI-MS (m/z): Calculated for:C₂₀H₂₈ClN₃O₅S: 457.97, observed mass; 456.05 (M−H); HPLC purity: 97.7%.

Production Example 67a: Synthesis of(S)—N-(1-(3-(cyclopropylmethoxy)-4-(trifluoromethyl) phenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

Step-1: Synthesis of3-hydroxy-N-methoxy-N-methyl-4-(trifluoromethyl)benzamide (2-b)

To a stirred solution of compound 1-b (3.5 g, 16.9 mmol) in DCM (35 mL),was added triethylamine (4.72 mL, 33.9 mmol) followed by addition ofN,O-dimethylhydroxylamine.HCl (1.98 g, 20.3 mmol) and stirred at roomtemperature for 20 min. EDC.HCl (4.88 g, 25.4 mmol) was added at 0° C.and the reaction mixture was stirred at room temperature for 30 min(reaction deemed complete by TLC). The reaction mixture was quenchedwith NaHCO₃ solution and extracted with DCM. The combined organic layerwas washed with brine, dried over anhydrous Na₂SO₄ and concentratedunder reduced pressure to afford compound 2-b (2.0 g, crude). LCMS:250.15 (M+1).

Step-2: Synthesis of3-(cyclopropylmethoxy)-N-methoxy-N-methyl-4-(trifluoromethyl) benzamide(4-b)

To a stirred solution of compound 2-b (2.0 g, 8.03 mmol) in DMF (20 mL),was added K₂CO₃ (2.21 g, 16.0 mmol) followed by(bromomethyl)cyclopropane (0.9 mL, 9.63 mmol) and the reaction mixturerefluxed for 5 h (reaction deemed complete by TLC). The reaction mixturewas quenched with water and extracted with EtOAc. The combined organiclayer was washed with brine, dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The residue was purified by columnchromatography using 25-30% EtOAc/hexane to afford compound 4-b (1.0 g,40%). LCMS: 304.05 (M+1).

Step-3: Synthesis of 1-(3-(cyclopropylmethoxy)-4-(trifluoromethyl)phenyl) ethan-1-one (6-b)

To a stirred solution of compound 4-b (0.7 g, 2.31 mmol) in dry THF (15mL) was added methyl magnesium bromide (3.0 M in THF, 1.5 mL, 4.62 mmol)at −10° C. The resultant mixture was stirred at room temperature for 12h (reaction deemed complete by TLC). The reaction mixture was quenchedwith NH₄Cl solution and diluted with EtOAc. The reaction mixture wasfiltered through a pad of Celite and extracted with EtOAc, washed withbrine, dried over anhydrous Na₂SO₄ and evaporated under reducedpressure. The residue was purified by column chromatography using 10%EtOAc/hexane to afford compound 6-b (0.4 g, 67.4%). LCMS: 259.20 (M+1).

Step-4: Synthesis of(S)—N-(1-(3-(cyclopropylmethoxy)-4-(trifluoromethyl)phenyl)ethylidene)-2-methylpropane-2-sulfinamide(8-b)

To a stirred solution of compound 6-b (0.4 g, 1.55 mmol) and compound7-b (0.3 g, 2.48 mmol) in dry toluene (10 mL) was added Ti(O^(i)Pr)₄(1.32 g, 4.65 mmol). The resultant mixture was heated at reflux for 16 h(reaction deemed complete by TLC). The reaction mixture was filteredthrough a pad of Celite, the filtrate was diluted with water andextracted with EtOAc. The combined organic layer was washed with brine,dried over anhydrous Na₂SO₄ and evaporated under reduced pressure. Theresidue was purified by column chromatography using 10% EtOAc/hexane toafford compound 8-b (0.33 g, 60%). LCMS: 362.1 (M+1).

Step-5: Synthesis of(S)—N—((S)-1-(3-(cyclopropylmethoxy)-4-(trifluoromethyl) phenyl)ethyl)-2-methylpropane-2-sulfinamide (9-b)

To a stirred solution of DIBAL-H (1M solution in toluene, 1.85 mL, 2.77mmol) in dry toluene (5 mL), was added a solution of compound 8-b (0.33g, 0.92 mmol) in toluene (5 mL) dropwise at −78° C. The resultantmixture was stirred at same temperature for 3 h (reaction deemedcomplete by TLC). The reaction mixture was quenched with NH₄Cl solutionand diluted with EtOAc. The reaction mixture was filtered through a padof Celite and extracted with EtOAc, washed with brine, dried overanhydrous Na₂SO₄ and evaporated under reduced pressure to affordcompound 9-b (0.33 g, crude). LCMS: 364.15 (M+1).

Step-6: Synthesis of (S)-1-(3-(cyclopropylmethoxy)-4-(trifluoromethyl)phenyl) ethan-1-amine hydrochloride (10-b)

To a stirred solution of compound 9-b (0.33 g, 0.91 mmol) in MeOH (8mL), was added 4M HCl in dioxane (0.5 mL) and the resulting mixturestirred at room temperature for 3 h (reaction deemed complete by TLC).The reaction mixture was concentrated and the residue was purified bytrituration with diethyl ether to afford 10-b (0.14 g, 53%). LCMS: 261.1(M+1).

Step 7: Synthesis of(S)—N-(1-(3-(cyclopropylmethoxy)-4-(trifluoromethyl) phenyl)ethyl)-5-(1,3-dioxoisoindolin-2-yl)pentane-1-sulfonamide (12-b)

To a stirred solution of compound 10-b —HCl salt (0.14 g, 0.48 mmol) inCH₂Cl₂ (5 mL) was added triethylamine (0.24 mL, 2.42 mmol) at 0° C. andstirred. To this reaction mixture was added compound 11-b (0.22 g, 0.72mmol) in CH₂Cl₂ (5 mL) dropwise over 25 min at 0° C. and stirred at thesame temperature for 3 h. The reaction was monitored by TLC for completeconsumption of compound 11-b. The reaction mixture was quenched withwater (25 mL) and extracted with CH₂Cl₂ (75 mL×2). The combined organicextracts was washed with brine (25 mL), dried over anhydrous sodiumsulfate and concentrated under reduced pressure. The crude was purifiedby silica gel flash chromatography using 15-20% EtOAc/hexanes to afford12-b as an off-white solid (0.17 g, 67%). LCMS: 539.15 (M+1).

Step 8: Synthesis of(S)-5-amino-N-(1-(3-(cyclopropylmethoxy)-4-(trifluoromethyl) phenyl)ethyl) pentane-1-sulfonamide (13-b)

To a stirred solution of compound 12-b (0.17 g, 0.32 mmol) in methanol(5 mL) was added hydrazine hydrate (99%, 0.19 mL, 1.62 mmol) at 0° C.Then ice bath was removed and the reaction mixture was warmed to roomtemperature and stirred for 3 h. The reaction was monitored by TLC.After completion, methanol was removed from reaction mixture underreduced pressure. The crude material was dissolved in 2N HCl (6 mL) andwashed with diethyl ether (10 mL×3). The aqueous layer was basified withaq. ammonia (pH=˜8) and extracted with ethyl acetate (10 mL×2). Theorganic extracts was dried over anhydrous sodium sulfate andconcentrated under reduced pressure to afford 13-b as a sticky mass(0.11 g, crude). This product was carried forward to the next stepwithout purification. LCMS: 409.2 (M+1).

Step 9: Synthesis of ethyl(S)-(5-(N-(1-(3-(cyclopropylmethoxy)-4-(trifluoromethyl) phenyl) ethyl)sulfamoyl) pentyl) glycinate (15-b)

To a solution of compound 13-b (0.11 g, 0.28 mmol) in ethanol (5 mL),was added ethyl 2-oxoacetate (50% solution in toluene, 0.03 mL, 0.31mmol) and stirred at room temperature for 1 h. To this was added asolution of NaCNBH₃ (20 mg, 0.33 mmol) in ethanol (3 mL; containing 1drop of AcOH) dropwise over 10 min at room temperature and reactionmixture was further stirred for 5 h. The reaction was monitored by TLCfor complete consumption of compound 13-b. Ethanol was removed underreduced pressure. The residue was taken in saturated NaHCO₃ solution (25mL) and extracted with EtOAc (25 mL×2). The organic extract was washedwith brine (25 mL) respectively then dried over anhydrous sodium sulfateand concentrated under reduced pressure to afford 15-b (0.04 g, crude).It was carried forward to the next step without purification. LCMS:495.1 (M+1).

Step 10: Synthesis of(S)—N-(1-(3-(cyclopropylmethoxy)-4-(trifluoromethyl) phenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide (ProductionExample 67a)

To a stirred solution of compound 15-b (0.04 g, 0.08 mmol) in AcOH (2mL) was added KOCN (14 mg, 0.16 mmol) and the reaction mixture wasstirred at room temperature for 16 h then heated at 60° C. for 12 h. Theprogress of the reaction was monitored by TLC. Reaction mixture wasconcentrated under reduced pressure and residue was taken in saturatedsolution of NaHCO₃ solution (15 mL) and extracted with EtOAc (15 mL×2).The organic extract was washed with water (15 mL×2) and brine (20 mL)respectively then dried over anhydrous sodium sulfate and concentratedunder reduced pressure to dryness. The residue was purified bycombiflash chromatography (eluting with 3-4% MeOH in DCM) to affordProduction Example 67a as a white solid (40 mg, crude). 40 mg crudecompound subject to prep HPLC purification which afforded 4 mg ofProduction Example 67a compound with 76% HPLC purity.

Production Example 68a: Synthesis of(S)—N-(1-(3-(cyclopropylmethoxy)-4-methylphenyl) ethyl)-5-(2,4-dioxoimidazolidin-1-yl) pentane-1-sulfonamide

Step-1: Synthesis of 1-(3-(cyclopropylmethoxy)-4-methylphenyl)ethan-1-one (2-c)

To a stirred solution of compound 1-c (3.0 g, 20.0 mmol) in DMF (50 mL),was added K₂CO₃ (8.2 g, 60.0 mmol) followed by addition of(bromomethyl)cyclopropane (3.2 g, 24 mmol) and the reaction mixture wasrefluxed for 4 h (reaction deemed complete by TLC). The reaction mixturewas quenched with water and extracted with EtOAc. The combined organiclayer was washed with brine, dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure to afford 3-c (3.2 g, crude). LCMS:205.05 (M+1).

Step-2: Synthesis of(S)—N-(1-(3-(cyclopropylmethoxy)-4-methylphenyl)ethylidene)-2-methylpropane-2-sulfinamide(5-c)

To a stirred solution of compound 3-c (3.2 g, 15.6 mmol) and compound4-c (2.8 g, 23.5 mmol) in dry toluene (150 mL) was added Ti(O^(i)Pr)₄(8.9 g, 31.3 mmol). The reaction mixture was diluted with EtOAc andquenched with water, filtered through a pad of Celite. The filtrate wasdiluted with water and extracted with EtOAc. The combined organic layerwas washed with brine, dried over anhydrous Na₂SO₄ and evaporated underreduced pressure. The residue was purified by column chromatographyusing 10% EtOAc/hexane to afford compound 5-c (2.8 g, 62%). LCMS: 308.15(M+1).

Step-3: Synthesis of(S)—N—((S)-1-(3-(cyclopropylmethoxy)-4-methylphenyl) ethyl)-2-methylpropane-2-sulfinamide (6-c)

To a stirred solution of DIBAL-H (1M solution in toluene, 2.7 mL, 27.3mmol) in dry toluene (15 mL), was added a solution of compound 5-c (2.8g, 9.1 mmol) in toluene (10 mL) dropwise at −78° C. The resultantmixture was stirred at same temperature for 3 h (reaction deemedcomplete by TLC). The reaction mixture was quenched with NH₄Cl solutionand diluted with EtOAc. The reaction mixture was filtered through a padof Celite and extracted with EtOAc, washed with brine, dried overanhydrous Na₂SO₄ and evaporated under reduced pressure. The residue waspurified by column chromatography using 10% EtOAc/hexane to affordcompound 6-c (2.0 g, 71%). LCMS: 310 (M+1).

Step-4: Synthesis of (S)-1-(3-(cyclopropylmethoxy)-4-methylphenyl)ethan-1-amine hydrochloride (7-c)

To a stirred solution of compound 6-c (2.0 g, 6.47 mmol) in MeOH (20mL), was added 4M HCl in dioxane (3.2 mL, 12.9 mmol) and the resultingmixture stirred at room temperature for 3 h (reaction deemed complete byTLC). The reaction mixture was concentrated and the residue was purifiedby trituration with diethyl ether to afford 7-c (0.9 g, 61%). LCMS:207.85 (M+1).

¹H NMR (400 MHz, DMSO-d₆) δ 8.60 (s, 3H), 7.10-7.07 (d, J=8.9 Hz, 2H),6.98-6.88 (d, J=7.5 Hz, 1H), 4.15-4.12 (m, 1H), 4.05-4.0 (m, 1H),3.83-3.85 (m, 2H), 2.12 (s, 3H), 1.46-1.35 (m, 3H), 0.62-0.50 (m, 2H),0.33-0.23 (m, 2H).

Step 5: Synthesis of (S)—N-(1-(3-(cyclopropylmethoxy)-4-methylphenyl)ethyl)-5-(1,3-dioxoisoindolin-2-yl) pentane-1-sulfonamide (9-c)

To a stirred solution of compound 7-c —HCl salt (0.3 g, 1.23 mmol) inCH₂Cl₂ (5 mL) was added triethylamine (0.68 mL, 4.89 mmol) at 0° C. andstirred. To this reaction mixture was added compound 8-c (0.47 g, 1.48mmol) in CH₂Cl₂ (5 mL) dropwise over 25 min at 0° C. and stirred at thesame temperature for 3 h. The reaction was monitored by TLC for completeconsumption of compound 7-c. The reaction mixture was quenched withwater (50 mL) and extracted with CH₂Cl₂ (50 mL×2). The combined organiclayer extract was washed with water (50 mL×2) and brine (50 mL), driedover anhydrous sodium sulfate and concentrated under reduced pressure.The crude was purified by silica gel flash chromatography using 15-20%EtOAc/hexanes to afford 9-c as an off-white solid (0.42 g, 70%). LCMS:485.2 (M+1).

Step 6: Synthesis of(S)-5-amino-N-(1-(3-(cyclopropylmethoxy)-4-methylphenyl) ethyl)pentane-1-sulfonamide (10-c)

To a stirred solution of compound 9-c (0.44 g, 0.9 mmol) in methanol (5mL) was added hydrazine hydrate (99%, 227 mL, 4.54 mmol) at 0° C. andstirred at room temperature for 3 h. Then ice bath was removed and thereaction mixture was warmed to room temperature and stirred for 5 h. Thereaction was monitored by TLC till the complete consumption of compound9-c. After completion, methanol was removed from reaction mixture underreduced pressure. The crude material was dissolved in 2N HCl (10 mL) andwashed with diethyl ether (10 mL×3). The aqueous layer was basified withaq. ammonia (pH=˜8) and extracted with ethyl acetate (10 mL×4). Theorganic extracts was dried over anhydrous sodium sulfate andconcentrated under reduced pressure to afford 10-c as a sticky mass(0.22 g, 68% yield). This product was carried forward to the next stepwithout purification. LCMS: 355.15 (M+1).

Step 7: Synthesis of ethyl(S)-(5-(N-(1-(3-(cyclopropylmethoxy)-4-methylphenyl) ethyl) sulfamoyl)pentyl)glycinate (11-c)

To a solution of compound 10-c (0.22 g, 0.62 mmol) in ethanol (5 mL) wasadded ethyl 2-oxoacetate (50% solution in toluene, 0.15 mL, 0.68 mmol)and stirred at room temperature for 1 h. To this was added a solution ofNaCNBH₃ (47 mg, 0.74 mmol) in ethanol (5 mL; containing 2 drops of AcOH)dropwise over 10 min at room temperature and reaction mixture wasfurther stirred for 3 h. The reaction was monitored by TLC for completeconsumption of compound 10-c. Ethanol was removed under reducedpressure. The residue was taken in saturated NaHCO₃ solution (10 mL) andextracted with EtOAc (10 mL×3). The organic extract was washed withwater (5 mL×3) and brine (10 mL) respectively then dried over anhydroussodium sulfate and concentrated under reduced pressure to afford 11-c(0.3 g, crude). It was carried forward to the next step withoutpurification. LCMS: 441.15 (M+1).

Step 8: Synthesis of (S)—N-(1-(3-(cyclopropylmethoxy)-4-methylphenyl)ethyl)-5-(2, 4-dioxoimidazolidin-1-yl) pentane-1-sulfonamide (ProductionExample 68a)

To a stirred solution of compound 11-c (0.3 g, 0.68 mmol) in AcOH (5 mL)was added KOCN (110 mg, 1.36 mmol) and the reaction mixture was stirredat room temperature for 16 h then heated at 60° C. for 6 h. The progressof the reaction was monitored by TLC. The reaction mixture wasconcentrated under reduced pressure and the residue treated withsaturated solution of NaHCO₃ solution (10 mL) and extracted with EtOAc(15 mL×2). The organic extract was washed with water (10 mL×2) and brine(10 mL) respectively then dried over anhydrous sodium sulfate andconcentrated under reduced pressure to dryness. The residue was purifiedby combiflash chromatography (eluting with 3-4% MeOH in DCM) to affordProduction Example 68a as a white solid (120 mg, 40%).

¹H NMR (400 MHz, DMSO-d₆) δ 10.69 (s, 1H), 7.62 (d, J=8.9 Hz, 1H), 7.06(d, J=7.5 Hz, 1H), 6.97 (d, J=1.7 Hz, 1H), 6.81 (dd, J=7.6, 1.6 Hz, 1H),4.40-4.28 (m, 1H), 3.89-3.80 (m, 4H), 3.11 (t, J=7.0 Hz, 2H), 2.72-2.62(m, 2H), 2.51-2.40 (m, 2H), 2.12 (s, 3H), 1.46-1.35 (m, 3H), 1.38-1.18(m, 2H), 1.11-1.05 (m, 2H), 0.98-0.95 (m, 1H), 0.62-0.50 (m, 2H),0.33-0.23 (m, 2H); ESI-MS (m/z): Calculated for: C₂₁H₃₁N₃O₅S: 437.56,observed mass; 436.1 (M−H); HPLC purity: 96.07%.

Production Example 69a: Synthesis of(S)—N-(1-(5-(cyclopropylmethoxy)-2-fluorophenyl) ethyl)-5-(2,4-dioxoimidazolidin-1-yl) pentane-1-sulfonamide

Step-1: Synthesis of1-(5-(cyclopropylmethoxy)-2-fluorophenyl)ethan-1-one (3-d)

To a stirred solution of 1-d (2.5 g, 16.2 mmol) in DMF (30 mL), wasadded K₂CO₃ (6.7 g, 48.7 mmol) followed by addition of(bromomethyl)cyclopropane (2.6 g, 19.4 mmol) and the reaction mixturewas refluxed for 5 h (reaction deemed complete by TLC). The reactionmixture was quenched with water and extracted with EtOAc. The combinedorganic layer was washed with brine, dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure to afford 3-d (2.6 g, crude). LCMS:209.0 (M+1).

Step-2: Synthesis ofN-(1-(5-(cyclopropylmethoxy)-2-fluorophenyl)ethylidene)pivalamide (5-d)

To a stirred solution of compound 3-d (2.06 g, 12.44 mmol) and compound4-d (2.25 g, 18.6 mmol) in dry toluene (50 mL) was added Ti(O^(i)Pr)₄(7.0 g, 24.8 mmol). The resultant mixture was heated at 90° C. for 16 h(reaction deemed complete by TLC). The reaction mixture was diluted withEtOAc and quenched with water, filtered through a pad of Celite. Thefiltrate was diluted with water and extracted with EtOAc. The combinedorganic layer was washed with brine, dried over anhydrous Na₂SO₄ andevaporated under reduced pressure. The residue was purified by columnchromatography using 10% EtOAc/hexane to afford compound 5-d (1.6 g,42%). LCMS: 292.05 (M+1).

Step-3: Synthesis of (S)—N-(1-(5-(cyclopropylmethoxy)-2-fluorophenyl)ethyl) pivalamide (6-d)

To a stirred solution of DIBAL-H (1M solution in toluene, 12.8 mL, 12.86mmol) in dry toluene (5 mL), was added a solution of compound 5-d (1.6g, 5.14 mmol) in toluene (10 mL) dropwise at −78° C. The resultantmixture was stirred at same temperature for 3 h (reaction deemedcomplete by TLC). The reaction mixture was quenched with NH₄Cl solutionand diluted with EtOAc. The reaction mixture was filtered through a padof Celite and extracted with EtOAc, washed with brine, dried overanhydrous Na₂SO₄ and evaporated under reduced pressure. The residue waspurified by column chromatography using 10% EtOAc/hexane to affordcompound 6-d (0.8 g, 50%). LCMS: 294.15 (M+1).

Step-4: Synthesis of (S)—N-(1-(5-(cyclopropylmethoxy)-2-fluorophenyl)ethyl) pivalamide (7-d)

To a stirred solution of compound 6-d (0.8 g, 2.55 mmol) in MeOH (5 mL),was added 4M HCl in dioxane (1.27 mL, 5.11 mmol) and the resultingmixture stirred at room temperature for 3 h (reaction deemed complete byTLC). The reaction mixture was concentrated and the residue was purifiedby trituration with diethyl ether to afford 7-d (0.4 g, 63%).

¹H NMR (400 MHz, DMSO-d₆) δ 8.60 (s, 3H), 7.10-7.09 (m, 2H), 6.98-6.94(m, 1H), 4.60-4.55 (m, 1H), 3.82 (s, 2H), 1.45 (s, 3H), 1.25-1.20 (m,2H), 0.61-0.52 (m, 2H), 0.35-0.27 (m, 2H).

Step 5: Synthesis of (S)—N-(1-(5-(cyclopropylmethoxy)-2-fluorophenyl)ethyl)-5-(1,3-dioxoisoindolin-2-yl) pentane-1-sulfonamide (9-d)

To a stirred solution of compound 7-d —HCl salt (0.4 g, 1.63 mmol) inCH₂Cl₂ (10 mL) was added triethylamine (0.68 mL, 4.89 mmol) at 0° C. andstirred. To this reaction mixture was added compound 8-d (0.56 g, 1.79mmol) in CH₂Cl₂ (5 mL) dropwise over 25 min at 0° C. and stirred at thesame temperature for 3 h. The reaction was monitored by TLC for completeconsumption of compound 7. The reaction mixture was quenched with water(50 mL) and extracted with CH₂Cl₂ (50 mL×2). The combined organicextracts were washed with water (50 mL×2) and brine (50 mL), dried overanhydrous sodium sulfate and concentrated under reduced pressure. Thecrude was purified by silica gel flash chromatography using 15-20%EtOAc/hexanes to afford 9-d as an off-white solid (0.25 g, 31%). LCMS:489.2 (M+1).

Step 6: Synthesis of(S)-5-amino-N-(1-(5-(cyclopropylmethoxy)-2-fluorophenyl) ethyl)pentane-1-sulfonamide (10-d)

To a stirred solution of compound 9-d (0.25 g, 0.51 mmol) in methanol (2mL) was added hydrazine hydrate (99%, 13 μL, 2.56 mmol) at 0° C. andstirred at room temperature for 3 h. Then ice bath was removed and thereaction mixture was warmed to room temperature and stirred for 5 h. Thereaction was monitored by TLC till the complete consumption of compound9-d. After completion, methanol was removed from reaction mixture underreduced pressure. The crude material was dissolved in 2N HCl (10 mL) andwashed with diethyl ether (10 mL×3). The aqueous layer was basified withaq. ammonia (pH=˜8) and extracted with ethyl acetate (10 mL×4). Theorganic extracts was dried over anhydrous sodium sulfate andconcentrated under reduced pressure to afford 10-d as a sticky mass(0.15 g, 84% yield). This product was carried forward to the next stepwithout purification. LCMS: 359.05 (M+1).

Step 7: Synthesis of ethyl(S)-(5-(N-(1-(5-(cyclopropylmethoxy)-2-fluorophenyl) ethyl) sulfamoyl)pentyl) glycinate (11-d)

To a solution of compound 10-d (0.15 g, 0.43 mmol) in ethanol (2 mL) wasadded ethyl 2-oxoacetate (50% solution in toluene, 0.1 mL, 0.47 mmol)and stirred at room temperature for 1 h. To this was added a solution ofNaCNBH₃ (33.1 mg, 0.51 mmol) in ethanol (5 mL; containing 2 drops ofAcOH) dropwise over 10 min at room temperature and reaction mixture wasfurther stirred for 4 h. The reaction was monitored by TLC for completeconsumption of compound 10-d. Ethanol was removed under reducedpressure. The residue was taken in saturated NaHCO₃ solution (10 mL) andextracted with EtOAc (10 mL×3). The organic extract was washed withwater (5 mL×3) and brine (10 mL) respectively then dried over anhydroussodium sulfate and concentrated under reduced pressure to afford 11-d(0.2 g, crude). It was carried forward to the next step withoutpurification. LCMS: 445.1 (M+1).

Step 8: Synthesis of (S)—N-(1-(5-(cyclopropylmethoxy)-2-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl) pentane-1-sulfonamide (ProductionExample 69a)

To a stirred solution of compound 11-d (0.2 g, 0.44 mmol) in AcOH (2 mL)was added KOCN (73 mg, 0.89 mmol) and the reaction mixture was stirredat room temperature for 16 h then heated at 60° C. for 6 h. The progressof the reaction was monitored by TLC. The reaction mixture wasconcentrated under reduced pressure and residue was taken in saturatedsolution of NaHCO₃ solution (10 mL) and extracted with EtOAc (15 mL×2).The organic extract was washed with water (10 mL×2) and brine (10 mL)respectively then dried over anhydrous sodium sulfate and concentratedunder reduced pressure to dryness. The residue was purified bycombiflash chromatography (eluting with 3-4% MeOH in DCM) to affordProduction Example 69a as a white solid (65 mg, 32% yield).

¹H NMR (400 MHz, DMSO-d₆) δ 10.70 (s, 1H), 7.77 (d, J=8.8 Hz, 1H),7.13-7.01 (m, 2H), 6.81-6.78 (m, 1H), 4.68-4.62 (m, 1H), 3.88 (s, 2H),3.78 (d, J=7.0 Hz, 2H), 3.14 (t, J=7.1 Hz, 2H), 2.88-2.80 (m, 1H),2.64-2.60 (m, 1H), 1.58-1.43 (m, 2H), 1.36-1.31 (m, 6H), 1.28-1.04 (m,2H), 0.61-0.52 (m, 2H), 0.35-0.27 (m, 2H); ESI-MS (m/z): Calculated for:C₂₀H₂₈FN₃O₅S: 441.52, observed mass; 440 (M−H); HPLC purity: 97.51%.

Production Example 70a: Synthesis of(S)—N-(1-(3-(cyclopropylmethoxy)-2-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

Step 1: Synthesis of1-(3-(cyclopropylmethoxy)-2-fluorophenyl)ethan-1-one (3-e)

To a stirred solution of 1-e (3.0 g, 19.4 mmol) in DMF (30 mL) was addedK₂CO₃ (8.0 g, 58.4 mmol) followed by addition of(bromomethyl)cyclopropane (2-e) (3.1 g, 23.3 mmol) and the reactionmixture was refluxed for 4 h (reaction deemed complete by TLC). Thereaction mixture was quenched with water and extracted with EtOAc. Thecombined organic layer was washed with brine, dried over anhydrousNa₂SO₄ and concentrated under reduced pressure to afford 3-e (3.4 g,crude).

Step 2: Synthesis of(S)—N-(1-(3-(cyclopropylmethyl)-2-fluorophenyl)ethylidene)-2-methylpropane-2-sulfinamide(5-e)

To a stirred solution of compound 3-e (3.4 g, 16.2 mmol) and compound4-e (2.95 g, 24.4 mmol) in dry toluene (50 mL) was added Ti(O^(i)Pr)₄(9.2 g, 32.5 mmol). The resultant mixture was heated at 90° C. for 16 h(reaction deemed complete by TLC). The reaction mixture was diluted withEtOAc and quenched with water, filtered through a pad of Celite. Thefiltrate was diluted with water and extracted with EtOAc. The combinedorganic layer was washed with brine, dried over anhydrous Na₂SO₄ andevaporated under reduced pressure. The residue was purified by columnchromatography using 10% EtOAc/hexane to afford compound 5-e (3.2 g,63%).

¹H NMR (400 MHz, DMSO-d₆) δ 7.12-7.08 (m, 1H), 7.05-7.01 (m, 2H),3.82-3.87 (m, 2H), 2.76 (s, 2H), 1.32 (s, 9H), 0.67-0.64 (m, 2H),0.38-0.36 (m, 2H).

Step 3: Synthesis of(S)—N—((S)-1-(3-(cyclopropylmethoxy)-2-fluorophenyl)ethyl)-2-methylpropane-2-sulfinamide (6-e)

To a stirred solution of DIBAL-H (1M solution in toluene, 30.0 mL, 30.8mmol) in dry toluene (10 mL), was added a solution of compound 5-e (3.2g, 10.2 mmol) in toluene (10 mL) dropwise at −78° C. The resultantmixture was stirred at same temperature for 3 h (reaction deemedcomplete by TLC). The reaction mixture was quenched with NH₄Cl solutionand diluted with EtOAc. The reaction mixture was filtered through a padof Celite and extracted with EtOAc, washed with brine, dried overanhydrous Na₂SO₄ and evaporated under reduced pressure. The residue waspurified by column chromatography using 10% EtOAc/hexane to affordcompound 6-e (1.8 g, 56%).

Step 4: Synthesis of (S)-1-(3-(cyclopropylmethoxy)-2-fluorophenyl)ethan-1-amine hydrochloride (7-e)

To a stirred solution of compound 6-e (1.8 g, 5.75 mmol) in MeOH (20mL), was added 4M HCl in dioxane (2.8 mL, 11.5 mmol) and the resultingmixture stirred at room temperature for 3 h (reaction deemed complete byTLC). The reaction mixture was concentrated and the residue was purifiedby trituration with diethyl ether to afford 7-e (0.9 g, 64%).

¹H NMR (400 MHz, DMSO-d₆) δ 8.62 (s, 3H), 7.21-7.08 (m, 3H), 4.61-4.48(m, 1H), 3.82-3.80 (m, 2H), 1.42-1.40 (m, 2H), 1.22-1.18 (m, 1H),0.60-0.56 (m, 2H), 0.38-0.26 (m, 2H).

Step 5: Synthesis of (S)—N-(1-(3-(cyclopropylmethoxy)-2-fluorophenyl)ethyl)-5-(1,3-dioxoisoindolin-2-yl)pentane-1-sulfonamide (9-e)

To a stirred solution of compound 7-e —HCl salt (0.3 g, 1.22 mmol) inCH₂Cl₂ (5 mL) was added triethylamine (0.5 mL, 3.67 mmol) at 0° C. andstirred. To this reaction mixture was added compound 8-e (0.46 g, 1.46mmol) in CH₂Cl₂ (5 mL) dropwise over 25 min at 0° C. and stirred at thesame temperature for 3 h. The reaction was monitored by TLC for completeconsumption of compound 7-e. The reaction mixture was quenched withwater (50 mL) and extracted with CH₂Cl₂ (50 mL×2). The combined organicextracts were washed with water (50 mL×2) and brine (50 mL), dried overanhydrous sodium sulfate and concentrated under reduced pressure. Thecrude was purified by silica gel flash chromatography using 15-20%EtOAc/hexanes to afford 9-e as an off-white solid (0.43 g, 72%). LCMS:489.15 (M+1).

Step 6: Synthesis of(S)-5-amino-N-(1-(3-(cyclopropylmethoxy)-2-fluorophenyl) ethyl)pentane-1-sulfonamide (10-e)

To a stirred solution of compound 9-e (0.42 g, 0.86 mmol) in methanol (5mL), was added hydrazine hydrate (99%, 215 mg, 4.30 mmol) at 0° C. andstirred at room temperature for 3 h. Then ice bath was removed and thereaction mixture was warmed to room temperature and stirred for 5 h. Thereaction was monitored by TLC till the complete consumption of compound9-e. After completion, methanol was removed from reaction mixture underreduced pressure. The crude material was dissolved in 2N HCl (10 mL) andwashed with diethyl ether (25 mL×3). The aqueous layer was basified withaq. ammonia (pH=˜8) and extracted with ethyl acetate (20 mL×2). Theorganic extracts was dried over anhydrous sodium sulfate andconcentrated under reduced pressure to afford 10-e as a sticky mass(0.15 g, 84% yield). This product was carried forward to the next stepwithout purification. LCMS: 359 (M+1).

Step 7: Synthesis of ethyl(S)-(5-(N-(1-(3-(cyclopropylmethoxy)-2-fluorophenyl) ethyl) sulfamoyl)pentyl) glycinate (11-e)

To a solution of compound 10-e (0.26 g, 0.72 mmol) in ethanol (4 mL) wasadded ethyl 2-oxoacetate (50% solution in toluene, 163 μL, 0.79 mmol)and stirred at room temperature for 1 h. To this was added a solution ofNaCNBH₄ (56.7 mg, 0.87 mmol) in ethanol (5 mL; containing 2 drops ofAcOH) dropwise over 10 min at room temperature and reaction mixture wasfurther stirred for 4 h. The reaction was monitored by TLC for completeconsumption of compound 10-e. Ethanol was removed under reducedpressure. The residue was taken in saturated NaHCO₃ solution (25 mL) andextracted with EtOAc (25 mL×3). The organic extract was washed withwater (20 mL×2) and brine (20 mL) respectively then dried over anhydroussodium sulfate and concentrated under reduced pressure to afford 11-e(0.3 g, crude). It was carried forward to the next step withoutpurification. LCMS: 445.15 (M+1).

Step 8: Synthesis of (S)—N-(1-(3-(cyclopropylmethoxy)-2-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide (ProductionExample 70a)

To a stirred solution of compound 11-e (0.3 g, 0.67 mmol) in AcOH (2 mL)was added KOCN (0.1 g, 1.34 mmol) and the reaction mixture was stirredat room temperature for 16 h then heated at 60° C. for 6 h. The progressof the reaction was monitored by TLC. Reaction mixture was concentratedunder reduced pressure and residue was taken in saturated solution ofNaHCO₃ solution (15 mL) and extracted with EtOAc (15 mL×2). The organicextract was washed with water (15 mL×2) and brine (15 mL) respectivelythen dried over anhydrous sodium sulfate and concentrated under reducedpressure to dryness. The residue was purified by combiflashchromatography (eluting with 3-4% MeOH in DCM) to afford ProductionExample 70a as a white solid (0.123 mg, 41%).

¹H NMR (400 MHz, DMSO-d₆) δ 10.68 (s, 1H), 7.80 (d, J=8.5 Hz, 1H),7.13-6.94 (m, 3H), 4.71-4.65 (m, 1H), 3.89-3.77 (m, 4H), 3.13 (t, J=7.0Hz, 2H), 2.84-2.80 (m, 1H), 2.62-2.59 (m, 1H), 1.58-1.51 (m, 2H), 1.35(t, J=7.8 Hz, 5H), 1.28-1.02 (m, 3H), 0.56-0.52 (m, 2H), 0.29-0.22 (m,2H); ESI-MS (m/z): Calculated for: C₂₀H₂₈FN₃O₅S: 441.52, observed mass;440.05 (M−H); HPLC purity: 99.08%.

Production Example 71a: Synthesis of(S)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-propoxyphenyl)ethyl) pentane-1-sulfonamide

Step 1: Synthesis of 1-(4-fluoro-3-propoxyphenyl) ethan-1-one (3-f)

To a stirred solution of 1-f (3.0 g, 19.4 mmol) in DMF (50 mL) was addedK₂CO₃ (8.0 g, 58.4 mmol) followed by addition of 1-bromopropane (2-f)(4.75 g, 38.9 mmol) and the reaction mixture was refluxed for 4 h(reaction deemed complete by TLC). The reaction mixture was quenchedwith water and extracted with EtOAc. The combined organic layer waswashed with brine, dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to afford 3-f (4.0 g, crude).

¹H NMR (400 MHz, DMSO-d₆) δ 7.65-7.57 (m, 1H), 7.52-7.48 (m, 1H), 7.12(t, J=53 Hz, 1H), 4.06-4.03 (m, 2H), 2.66 (s, 3H), 1.92-1.82 (m, 2H).1.07-1.04 (m, 3H).

Step 2: Synthesis of(S)—N-(1-(4-fluoro-3-propoxyphenyl)ethylidene)-2-methylpropane-2-sulfinamide(5-f)

To a stirred solution of compound 3-f (4.0 g, 20.4 mmol) and compound4-f (3.95 g, 51.0 mmol) in dry toluene (40 mL) was added Ti(O^(i)Pr)₄(14.4 g, 32.6 mmol). The resultant mixture was heated at 90° C. for 16 h(reaction deemed complete by TLC). The reaction mixture was diluted withEtOAc and quenched with water, filtered through a pad of Celite. Thefiltrate was diluted with water and extracted with EtOAc. The combinedorganic layer was washed with brine, dried over anhydrous Na₂SO₄ andevaporated under reduced pressure. The residue was purified by columnchromatography to afford compound 5-f (5.82 g, crude). LCMS: 300.05(M+1).

Step 3: Synthesis of (S)—N—((S)-1-(4-fluoro-3-propoxyphenyl)ethyl)-2-methylpropane-2-sulfinamide (6-f)

To a stirred solution of DIBAL-H (1M solution in toluene, 25 mL, 25.0mmol) in dry toluene (10 mL), was added a solution of compound 5-f (3.0g, 10.0 mmol) in toluene (10 mL) dropwise at −78° C. The resultantmixture was stirred at same temperature for 3 h (reaction deemedcomplete by TLC). The reaction mixture was quenched with NH₄Cl solutionand diluted with EtOAc. The reaction mixture was filtered through a padof Celite and extracted with EtOAc, washed with brine, dried overanhydrous Na₂SO₄ and evaporated under reduced pressure. The residue waspurified by column chromatography using 10% EtOAc/hexane to affordcompound 6-f (1.6 g, 8.53%). LCMS: 302.15 (M+1).

Step 4: Synthesis of (S)-1-(4-fluoro-3-propoxyphenyl) ethan-1-aminehydrochloride (7-f)

To a stirred solution of compound 6-f (1.6 g, 5.31 mmol) in MeOH (5 mL),was added 4M HCl in dioxane (1.5 mL, 5.84 mmol) and the resultingmixture stirred at room temperature for 3 h (reaction deemed complete byTLC). The reaction mixture was concentrated and the residue was purifiedby trituration with diethyl ether to afford 7-f (0.7 g, 57%).

Step 5: Synthesis of(S)-5-(1,3-dioxoisoindolin-2-yl)-N-(1-(4-fluoro-3-propoxyphenyl)ethyl)pentane-1-sulfonamide (9-f)

To a stirred solution of compound 7-f —HCl salt (0.7 g, 3.0 mmol) inCH₂Cl₂ (5 mL) was added triethylamine (2.0 mL, 15.0 mmol) at 0° C. andstirred. To this reaction mixture was added compound 8-f (1.4 g, 4.5mmol) in CH₂Cl₂ (5 mL) dropwise over 25 min at 0° C. and stirred at thesame temperature for 3 h. The reaction was monitored by TLC for completeconsumption of compound 7-f. The reaction mixture was quenched withwater (50 mL) and extracted with CH₂Cl₂ (50 mL×2). The combined organicextracts were washed with brine (25 mL), dried over anhydrous sodiumsulfate and concentrated under reduced pressure. The crude was purifiedby silica gel flash chromatography using 15-20% EtOAc/hexanes to afford9-f as an off-white solid (1.4 g, 98%). LCMS: 494.2 (M−18).

Step 6: Synthesis of (S)-5-amino-N-(1-(4-fluoro-3-propoxyphenyl) ethyl)pentane-1-sulfonamide (10-f)

To a stirred solution of compound 8-f (1.5 g, 3.15 mmol) in methanol (5mL), was added hydrazine hydrate (99%, 0.78 mL, 15.5 mmol) at 0° C. andstirred at room temperature for 3 h. The ice bath was removed and thereaction mixture was warmed to room temperature and stirred for 5 h. Thereaction was monitored by TLC. After completion, methanol was removedfrom reaction mixture under reduced pressure. The crude material wasdissolved in 2N HCl (10 mL) and washed with diethyl ether (10 mL×3). Theaqueous layer was basified with aq. ammonia (pH=˜8) and extracted withethyl acetate (10 mL×4). The organic extracts was dried over anhydroussodium sulfate and concentrated under reduced pressure to afford 10-f asa sticky mass (1.0 g, 91.7% yield). This product was carried forward tothe next step without purification. LCMS: 347.05 (M+1).

Step 7: Synthesis of ethyl (S)-(5-(N-(1-(4-fluoro-3-propoxyphenyl)ethyl) sulfamoyl) pentyl) glycinate (11-f)

To a solution of compound 10-f (1.0 g, 2.89 mmol) in methanol (10 mL)was added ethyl 2-oxoacetate (50% solution in toluene, 0.3 mL, 3.17mmol) and stirred at room temperature for 1 h. To this was added asolution of NaCNBH₃ (0.21 mg, 3.46 mmol) in ethanol (5 mL; containing 2drops of AcOH) dropwise over 10 min at room temperature and reactionmixture was further stirred for 4 h. The reaction was monitored by TLCfor complete consumption of compound 10-f. Ethanol was removed underreduced pressure. The residue was taken in saturated NaHCO₃ solution (25mL) and extracted with EtOAc (25 mL×3). The organic extract was washedwith water (25 mL×3) and brine (25 mL) respectively then dried overanhydrous sodium sulfate and concentrated under reduced pressure toafford 11-f (0.5 g, crude). It was carried forward to the next stepwithout purification. LCMS: 433.1 (M+1).

Step 8: Synthesis of(S)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-propoxyphenyl)ethyl) pentane-1-sulfonamide (Production Example 71a)

To a stirred solution of compound 11-f (0.5 g, 1.16 mmol) in AcOH (4 mL)was added KOCN (0.18 g, 2.32 mmol) and the reaction mixture was stirredat room temperature for 16 h then heated at 60° C. for 6 h. The progressof the reaction was monitored by TLC. Reaction mixture was concentratedunder reduced pressure and residue was taken in saturated solution ofNaHCO₃ solution (50 mL) and extracted with EtOAc (50 mL×2). The organicextract was washed with water (50 mL×2) and brine (10 mL) respectivelythen dried over anhydrous sodium sulfate and concentrated under reducedpressure to dryness. The residue was purified by combiflashchromatography (eluting with 3-4% MeOH in DCM) to afford ProductionExample 71a as a white solid (30 mg, 6.48% yield).

¹H NMR (400 MHz, DMSO-d₆) δ 10.71-10.66 (m, 1H), 7.67 (d, J=8.8 Hz, 1H),7.25-7.09 (m, 2H), 6.91 (t, J=5.8 Hz, 1H), 4.40 (p, J=7.1 Hz, 1H), 3.99(t, J=6.6 Hz, 2H), 3.87 (s, 3H), 3.13 (t, J=7.1 Hz, 2H), 2.78-2.70 (m,1H), 2.62-2.52 (m, 1H), 1.82-1.68 (m, 3H), 1.58-1.40 (m, 3H), 1.40-0.94(m, 7H); ESI-MS (m/z): Calculated for: C₂₀H₂₈FN₃O₅S: 441.52, observedmass; 440 (M−H); HPLC purity: 97.51%.

Production Example 72a: Synthesis of(S)—N-(1-(3-(allyloxy)-4-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

Step 1: Synthesis of 1-(3-(allyloxy)-4-fluorophenyl)ethan-1-one (3-g)

To a stirred solution of 1-g (2.0 g, 12.9 mmol) in DMF (10 mL) was addedK₂CO₃ (5.37 g, 38.9 mmol) followed by addition of 3-bromoprop-1-ene(3-g) (3.14 g, 25.9 mmol) and the reaction mixture was refluxed for 2 h(reaction deemed complete by TLC). The reaction mixture was quenchedwith water and extracted with EtOAc. The combined organic layer waswashed with brine, dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to afford 3-g (1.8 g, crude). LCMS: 195.05 (M+1).

Step 2: Synthesis of(S)—N-(1-(3-(allyloxy)-4-fluorophenyl)ethylidene)-2-methylpropane-2-sulfinamide(5-g)

To a stirred solution of compound 3-g (1.8 g, 9.27 mmol) and compound4-g (1.79 g, 14.8 mmol) in dry toluene (20 mL) was added Ti(O^(i)Pr)₄(6.58 g, 23.1 mmol). The resultant mixture was heated at 90° C. for 16 h(reaction deemed complete by TLC). The reaction mixture was diluted withEtOAc and quenched with water, filtered through a pad of Celite. Thefiltrate was diluted with water and extracted with EtOAc. The combinedorganic layer was washed with brine, dried over anhydrous Na₂SO₄ andevaporated under reduced pressure. The residue was purified by columnchromatography to afford compound 5-g (2.7 g, crude). LCMS: 298.10(M+1).

Step 3: Synthesis of (S)—N—((S)-1-(3-(allyloxy)-4-fluorophenyl)ethyl)-2-methylpropane-2-sulfinamide (6-g)

To a stirred solution of DIBAL-H (1M solution in toluene, 1.94 mL, 10.9mmol) in dry toluene (5 mL), was added a solution of compound 5-g (2.7g, 9.09 mmol) in toluene (25 mL) dropwise at −78° C. The resultantmixture was stirred at same temperature for 3 h (reaction deemedcomplete by TLC). The reaction mixture was quenched with NH₄Cl solutionand diluted with EtOAc. The reaction mixture was filtered through a padof Celite and extracted with EtOAc, washed with brine, dried overanhydrous Na₂SO₄ and evaporated under reduced pressure. The residue waspurified by column chromatography using 10% EtOAc/hexane to affordcompound 6-g (2.0 g, 73.5%). LCMS: 300.1 (M+1).

Step 4: Synthesis of (S)-1-(3-(allyloxy)-4-fluorophenyl)ethan-1-aminehydrochloride (7-g)

To a stirred solution of compound 6-g (2.0 g, 6.68 mmol) in MeOH (20mL), was added 4M HCl in dioxane (2.0 mL, 7.35 mmol) and the resultingmixture stirred at room temperature for 3 h (reaction deemed complete byTLC). The reaction mixture was concentrated and the residue was purifiedby trituration with diethyl ether to afford 7-g (0.65 g, 50%).

¹H NMR (400 MHz, DMSO-d₆) δ 7.07-7.01 (m, 2H), 6.89-6.87 (m, 1H),6.12-6.02 (m, 1H), 5.45 (d, J=13 Hz, 1H), 5.30 (d, J=13 Hz, 1H), 4.63(s, 2H), 2.97 (s, 3H).

Step 5: Synthesis of (S)—N-(1-(3-(allyloxy)-4-fluorophenyl)ethyl)-5-(1,3-dioxoisoindolin-2-yl)pentane-1-sulfonamide (9-g)

To a stirred solution of compound 7-g —HCl salt (0.65 g, 3.31 mmol) inCH₂Cl₂ (5 mL) was added triethylamine (0.26 mL, 8.60 mmol) at 0° C. andstirred. To this reaction mixture was added compound 8-g (1.5 g, 4.97mmol) in CH₂Cl₂ (5 mL) dropwise over 25 min at 0° C. and stirred at thesame temperature for 3 h. The reaction was monitored by TLC for completeconsumption of compound 7-g. The reaction mixture was quenched withwater (50 mL) and extracted with CH₂Cl₂ (50 mL×2). The combined organicextracts were washed with brine (50 mL), dried over anhydrous sodiumsulfate and concentrated under reduced pressure. The crude was purifiedby silica gel flash chromatography using 15-20% EtOAc/hexanes to afford9-g as an off-white solid (0.8 g, 53.3%). LCMS: 475.05 (M+1).

Step 6: Synthesis of ((S)—N-(1-(3-(allyloxy)-4-fluorophenyl)ethyl)-5-aminopentane-1-sulfonamide (10-g)

To a stirred solution of compound 9-g (0.8 g, 1.68 mmol) in methanol (10mL) was added hydrazine hydrate (99%, 0.25 mL, 8.4 mmol) at 0° C. andstirred at room temperature for 3 h. Then ice bath was removed and thereaction mixture was warmed to room temperature and stirred for 5 h. Thereaction was monitored by TLC till the complete consumption of compound9-g. After completion, methanol was removed from reaction mixture underreduced pressure. The crude material was dissolved in 2N HCl (25 mL) andwashed with diethyl ether (25 mL×2). The aqueous layer was basified withaq. ammonia (pH=˜8) and extracted with ethyl acetate (25 mL×2). Theorganic extracts was dried over anhydrous sodium sulfate andconcentrated under reduced pressure to afford 10-g as sticky mass (0.59g, 86.6% yield). This product was carried forward to the next stepwithout purification. LCMS: 345.15 (M+1).

Step 7: Synthesis of ethyl (S)-(5-(N-(1-(3-(allyloxy)-4-fluorophenyl)ethyl) sulfamoyl) pentyl) glycinate (11-g)

To a solution of compound 10-g (0.5 g, 1.45 mmol) in ethanol (5 mL) wasadded ethyl 2-oxoacetate (50% solution in toluene, 0.16 mL, 1.59 mmol)and stirred at room temperature for 1 h. To this was added a solution ofNaCNBH₃ (0.10 mg, 1.74 mmol) in ethanol (5 mL; containing 2 drops ofAcOH) dropwise over 10 min at room temperature and the reaction mixturewas further stirred for 4 h. The reaction was monitored by TLC forcomplete consumption of compound 10-g. Ethanol was removed under reducedpressure. The residue was taken in saturated NaHCO₃ solution (15 mL) andextracted with EtOAc (20 mL×2). The organic extract was washed withwater (20 mL) and brine (15 mL) respectively then dried over anhydroussodium sulfate and concentrated under reduced pressure to afford 11-g(0.5 g, crude) which was carried forward to the next step withoutpurification. LCMS: 431.15 (M+1).

Step 8: Synthesis of (S)—N-(1-(3-(allyloxy)-4-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide (ProductionExample 72a)

To a stirred solution of compound 11-g (0.5 g, 1.16 mmol) in AcOH (4 mL)was added KOCN (0.18 g, 2.32 mmol) and the reaction mixture was stirredat room temperature for 16 h then heated at 60° C. for 6 h. The progressof the reaction was monitored by TLC. Reaction mixture was concentratedunder reduced pressure and residue was taken in saturated solution ofNaHCO₃ solution (50 mL) and extracted with EtOAc (50 mL×2). The organicextract was washed with water (50 mL) and brine (15 mL) respectivelythen dried over anhydrous sodium sulfate and concentrated under reducedpressure to dryness. The residue was purified by combiflashchromatography (eluting with 3-4% MeOH in DCM) to afford ProductionExample 72a as a white solid (25 mg, 5% yield).

¹H NMR (400 MHz, DMSO-d₆) δ 10.68 (s, 1H), 7.67 (d, J=8.8 Hz, 1H),7.27-7.11 (m, 2H), 6.98-6.90 (m, 1H), 6.05-6.0 (m, 1H), 5.43 (dd,J=17.2, 1.8 Hz, 1H), 5.29 (d, J=10.5 Hz, 1H), 4.63 (d, J=5.4 Hz, 2H),4.40 (p, J=7.1 Hz, 1H), 3.87 (s, 2H), 3.13 (t, J=7.1 Hz, 2H), 2.77-2.75(m, 1H), 2.62-2.53 (m, 1H), 1.49-1.40 (m, 2H), 1.40-1.00 (m, 7H); ESI-MS(m/z): Calculated for: C₂₀H₂₆FN₃O₅S: 427.49, observed mass; 426 (M−H);HPLC purity: 99.9%.

Production Example 73a: Synthesis of(S)—N-(1-(3-(2-cyclopropylethoxy)-4-fluorophenyl) ethyl)-5-(2,4-dioxoimidazolidin-1-yl) pentane-1-sulfonamide

Step 1: Synthesis of1-(3-(2-cyclopropylethoxy)-4-fluorophenyl)ethan-1-one (3-h)

To a stirred solution of 1-h (2.6 g, 16.8 mmol) in DMF (25 mL) was addedK₂CO₃ (4.65 g, 33.7 mmol) followed by (2-bromoethyl)cyclopropane (3.0 g,20.2 mmol) and the reaction mixture was refluxed for 2 h (reactiondeemed complete by TLC). The reaction mixture was quenched with waterand extracted with EtOAc. The combined organic layer was washed withbrine, dried over anhydrous sodium sulfate and concentrated underreduced pressure to afford 3-h (3.5 g, crude). LCMS: 223.15 (M+1).

Step 2: Synthesis of(S)—N-(1-(3-(2-cyclopropylethoxy)-4-fluorophenyl)ethylidene)-2-methylpropane-2-sulfinamide(5-h)

To a stirred solution of compound 3-h (2.0 g, 9.00 mmol) and compound4-h (1.63 g, 13.5 mmol) in dry toluene (40 mL) was added Ti(O^(i)Pr)₄(5.33 mL, 18.0 mmol). The resultant mixture was heated at 90° C. for 16h (reaction deemed complete by TLC). The reaction mixture was dilutedwith EtOAc and quenched with water, filtered through a pad of Celite.The filtrate was diluted with water and extracted with EtOAc. Thecombined organic layer was washed with brine, dried over anhydroussodium sulfate and evaporated under reduced pressure The residue waspurified by column chromatography to afford compound 5-h (3.0 g, crude).LCMS: 326.0 (M+1).

Step 3: Synthesis of(S)—N—((S)-1-(3-(2-cyclopropylethoxy)-4-fluorophenyl)ethyl)-2-methylpropane-2-sulfinamide(6-h)

To a stirred solution of DIBAL-H (1M solution in toluene, 18.4 mL, 27.6mmol) in dry toluene (20 mL), was added a solution of compound 5-h (3.0g, 9.23 mmol) in toluene (15 mL) dropwise at −78° C. The resultantmixture was stirred at −78° C. for 3 h (reaction deemed complete byTLC). The reaction mixture was quenched with NH₄Cl solution and dilutedwith EtOAc. The reaction mixture was filtered through a pad of Celiteand extracted with EtOAc, washed with brine, dried over anhydrous sodiumsulfate and evaporated under reduced pressure. The residue was purifiedby column chromatography using 10% EtOAc/hexane to afford compound 6-h(1.3 g, 43.1%). LCMS: 328.2 (M+1).

Step 4: Synthesis of(S)-1-(3-(2-cyclopropylethoxy)-4-fluorophenyl)ethan-1-aminehydrochloride (7-h)

To a stirred solution of compound 6-h (1.3 g, 3.97 mmol) in dioxane (8mL) was added 4M HCl in dioxane (8 mL) and the resulting mixture stirredat room temperature for 3 h (reaction deemed complete by TLC). Thereaction mixture was concentrated and the residue was purified bytrituration with pentane to afford 7-h (0.9 g, 87.3%). LCMS: 224.05(M+1).

Step 5: Synthesis of(S)—N-(1-(3-(2-cyclopropylethoxy)-4-fluorophenyl)ethyl)-5-(1,3-dioxoisoindolin-2-yl)pentane-1-sulfonamide(9-h)

To a stirred solution of compound 7-h —HCl salt (0.63 g, 2.41 mmol) inDCM (5 mL) was added triethylamine (1.68 mL, 12. mmol) at 0° C. andstirred. To this reaction mixture was added compound 8-h (1.14 g, 3.62mmol) in DCM (5 mL) dropwise over 25 min at 0° C. and stirred at thesame temperature for 3 h. The reaction was monitored by TLC. Thereaction mixture was quenched with water (50 mL) and extracted with DCM(50 mL×2). The combined organic extracts were washed with brine (50 mL),dried over anhydrous sodium sulfate and concentrated under reducedpressure. The crude was purified by silica gel flash chromatographyusing 15-20% EtOAc/hexanes to afford 9-h as an off-white solid (0.52 g,43%). LCMS: 501.05 (M-1).

Step 6: Synthesis of(S)-5-amino-N-(1-(3-(2-cyclopropylethoxy)-4-fluorophenyl) ethyl)pentane-1-sulfonamide (10-h)

To a stirred solution of compound 9-h (0.5 g, 0.99 mmol) in MeOH (5 mL)was added hydrazine hydrate (99%, 0.24 mL, 4.98 mmol) at 0° C. and thenthe ice bath was removed. The reaction mixture was warmed to roomtemperature and stirred for 3 h. The reaction was monitored by TLC tillthe complete consumption of compound 9-h. After completion, MeOH wasremoved from reaction mixture under reduced pressure. The crude materialwas dissolved in 2N HCl (25 mL) and washed with diethyl ether (25 mL×2).The aqueous layer was basified with saturated aqueous ammonia (pH=˜8)and extracted with EtOAc (25 mL×2). The organic extracts were dried overanhydrous sodium sulfate and concentrated under reduced pressure toafford 10-h as a yellow sticky solid (0.33 g, crude). This product wascarried forward to the next step without purification. LCMS: 373.05(M+1).

Step 7: Synthesis of ethyl(S)-(5-(N-(1-(3-(2-cyclopropylethoxy)-4-fluorophenyl) ethyl) sulfamoyl)pentyl) glycinate (11-h)

To a solution of compound 10-h (0.33 g, 0.88 mmol) in ethanol (8 mL) wasadded ethyl 2-oxoacetate (50% solution in toluene, 99 μL, 0.97 mmol) andstirred at room temperature for 1 h. To this was added a solution ofNaCNBH₃ (67 mg, 1.04 mmol) and AcOH (2 drops) in ethanol (8 mL) addeddropwise over 10 min to the reaction mixture at room temperature andreaction mixture was further stirred for 4 h. The reaction was monitoredby TLC for complete consumption of compound 10-h. Ethanol was removedunder reduced pressure. The residue was taken in saturated NaHCO₃solution (15 mL) and extracted with EtOAc (20 mL×2). The organic extractwas washed with water (20 mL) and brine (15 mL) respectively then driedover anhydrous sodium sulfate and concentrated under reduced pressure toafford 11-h (0.34 g, crude) which was carried forward to the next stepwithout purification. LCMS: 459.15 (M+1).

Step 8: Synthesis of (S)—N-(1-(3-(2-cyclopropylethoxy)-4-fluorophenyl)ethyl)-5-(2, 4-dioxoimidazolidin-1-yl) pentane-1-sulfonamide (ProductionExample 73a)

To a stirred solution of compound 11-h (0.34 g, 0.74 mmol) in AcOH (5mL) was added KOCN (0.12 g, 1.48 mmol) and the reaction mixture wasstirred at room temperature for 16 h then heated at 60° C. for 6 h. Theprogress of the reaction was monitored by TLC. Reaction mixture wasconcentrated under reduced pressure and residue was taken in saturatedsolution of NaHCO₃ solution (50 mL) and extracted with EtOAc (50 mL×2).The organic extract was washed with water (50 mL) and brine (15 mL)respectively then dried over anhydrous sodium sulfate and concentratedunder reduced pressure to dryness. The residue was purified by flashcolumn chromatography (eluting with 3-4% MeOH:DCM) to afford ProductionExample 73a as a white solid (85 mg, 25% yield).

¹H NMR (400 MHz, CDCl₃) δ 7.17 (dd, J=8.4, 6.6 Hz, 1H), 6.73-6.60 (m,2H), 5.40-5.29 (m, 1H), 4.72-4.59 (m, 1H), 4.09 (t, J=6.4 Hz, 2H), 3.88(s, 2H), 3.33-3.32 (m, 2H), 2.77-2.60 (m, 1H), 2.69-2.60 (m, 1H),1.84-1.51 (m, 7H), 1.49-1.39 (m, 2H), 1.35-1.17 (m, 3H), 0.85 (t, J=6.4Hz, 1H), 0.64-0.51 (m, 2H), 0.18 (d, J=5.1 Hz, 2H); ESI-MS (m/z):Calculated for: C₂₁H₃₀F₃O₅S: 455.55, observed mass; 454.3 (M−H); HPLCpurity: 98.5%.

Production Example 74a: Synthesis of (R)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-hydroxyphenyl) ethyl)pentane-1-sulfonamide

Step 1: Synthesis of (R)-5-(2,4-dioxoimidazolidin-1-yl)-N-(1-(4-fluoro-3-hydroxyphenyl) ethyl)pentane-1-sulfonamide (Production Example 74a)

To a stirred solution of Production Example 1 (0.15 g, 0.34 mmol) inMeOH (2 mL), was added 6N HCl (3 mL) at 80° C. for 18 h. After completeaddition, the reaction mixture was stirred was diluted with EtOAc (50mL×2). The organic layer was separated and washed with water (50 mL),followed by brine (50 mL) and dried over anhydrous sodium sulfate,filtered and concentrated under reduced pressure to obtain the crude.The crude product was purified by silica gel column chromatography(60-120 mesh) using 1-3% MeOH/DCM to afford Production Example 74a as anoff-white solid (60 mg, 45.6% yield).

¹H NMR (400 MHz, DMSO-d₆) δ 10.69 (s, 1H), 9.82 (s, 1H), 7.65 (d, J=8.3Hz, 1H), 7.07 (dd, J=11.3, 8.3 Hz, 1H), 6.95 (dd, J=8.5, 2.2 Hz, 1H),6.77-6.75 (m, 2H), 4.36-4.30 (m, 1H), 3.88 (s, 2H), 3.15 (t, J=7.1 Hz,2H), 2.78-2.74 (m, 1H), 2.56-2.53 (m, 1H), 1.61-1.31 (m, 6H), 1.12-1.07(m, 2H); ESI-MS (m/z): Calculated for: C₁₆H₂₂FN₃O₅S: 387.43, observedmass; 385.95 (M−1); HPLC purity: 99.26%.

Production Example 26a: Synthesis of(R)—N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)-6-(2,4-dioxoimidazolidin-1-yl)hexane-1-sulfonamide

Step 1: Synthesis of 2-(6-bromohexyl)isoindoline-1,3-dione (3-i)

To a stirred solution of 1,6-dibromohexane (12.3 mL, 81.0 mmol) in DMF(10 mL), was added potassium phthalate (5.0 g, 27.0 mmol) portion-wiseover 30 min at room temperature. After complete addition, the reactionmixture was stirred at 90° C. for 18 h, then quenched with water (300mL) and extracted with diethyl ether (150 mL×2). The combined organicextracts were washed with water (100 mL×2), followed by brine (50 mL×2)and dried over anhydrous sodium sulfate, filtered and concentrated underreduced pressure. The resultant residue was purified by silica gelcolumn chromatography (60-120 mesh) using 5-10% EtOAc/hexanes to afford3-i as an off-white solid (6.3 g, 76% yield). LCMS: 310.95 (M+1).

Step 2: Synthesis of S-(6-(1,3-dioxoisoindolin-2-yl)hexyl) ethanethioate(4-i)

To a stirred solution of compound 3-i (6.39 g, 20.3 mol) in DMF (60 mL)was added potassium thioacetate (2.78 g, 34.3 mmol) portion wise andstirred for 20 min at room temperature. After complete addition, thereaction mixture was stirred at room temperature for 30 min. Reactionprogress was monitored by TLC; after completion the reaction mixture wasquenched with ice-cold water (250 mL) and stirred for 1 h. The resultantprecipitated was filtered, washed with water (100 mL) and dried in vacuoto afford 4-i as an off-white solid (5.8 g, 93.5%). LCMS: 306.20 (M+1).

Step 3: Synthesis of 6-(1,3-dioxoisoindolin-2-yl)hexane-1-sulfonylchloride (5-i)

2N HCl (3.63 mL) was added to a stirred solution of compound 4-i (2.0 g,6.55 mol) in acetonitrile (36 mL) at 0° C. This was followed byportion-wise addition of N-chlorosuccinimide (3.85 g, 28.8 mol) over 30min and the reaction mixture was warmed to room temperature and stirredfor 1 h. Reaction progress was monitored by TLC. Upon completeconsumption of 4-i, the reaction mixture was quenched with ice-coldwater (100 mL) and extracted with diethyl ether (100 mL×2). The combinedorganic extracts were washed with saturated sodium bicarbonate solution(50 mL), water (100 mL) and brine (50 mL) and dried over anhydroussodium sulfate and concentrated under reduced pressure to affordcompound 5-i as an off-white solid (1.81 g, 83.4% yield).

¹H NMR (400 MHz, CDCl₃): δ 7.90-7.80 (m, 2H), 7.77-7.67 (m, 2H),3.74-3.59 (m, 4H), 2.12-1.98 (m, 2H), 1.72 (p, J=7.2 Hz, 2H), 1.62-1.32(m, 4H).

Step 4: Synthesis of (R)—N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)-6-(1, 3-dioxoisoindolin-2-yl) hexane-1-sulfonamide (7-i)

To a stirred solution of compound 6-i —HCl salt (0.5 g, 2.39 mmol) inDCM (15 mL), was added triethylamine (0.9 mL, 6.33 mmol) at 0° C. andstirred. To this reaction mixture was added compound 6-i (1.18 mL, 3.58mmol) in DCM (10 mL) dropwise over 25 min at 0° C. and stirred at thesame temperature for 3 h. The reaction was monitored by TLC for completeconsumption of compound 6-i. The reaction mixture was quenched withwater (50 mL) and extracted with DCM (50 mL×2). The combined organicextracts were washed with water (50 mL×2) and brine (50 mL), dried overanhydrous sodium sulfate and concentrated under reduced pressure. Theresultant residue was purified by silica gel flash chromatography using15-20% EtOAc/hexanes to afford 7-i as an off-white solid (1.0 g, 84%).LCMS: 520.20 (M+18).

Step 5: Synthesis of(R)-6-amino-N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl) ethyl)hexane-1-sulfonamide (8-i)

To a stirred solution of compound 7-i (1.0 g, 1.99 mmol) in MeOH (10mL), was added hydrazine hydrate (99%, 0.5 mL, 9.96 mmol) at 0° C. andstirred at room temperature for 3 h. The ice bath was removed and thereaction mixture was warmed to room temperature and stirred for 5 h. Thereaction was monitored by TLC. After completion, MeOH was removed fromreaction mixture under reduced pressure. The crude material wasdissolved in 2N HCl (50 mL) and washed with diethyl ether (50 mL×3). Theaqueous layer was basified with aq. ammonia (pH=˜8) and extracted withEtOAc (50 mL×2). The organic extract was dried over anhydrous sodiumsulfate and concentrated under reduced pressure to afford 8-i as asticky mass (0.61 g, 82.3% yield). This material was carried forward tothe next step without purification. LCMS: 373.15 (M+1).

Step 6: Synthesis of ethyl(R)-(6-(N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl) ethyl)sulfamoyl)hexyl)glycinate (9-i)

To a solution of compound 8-i (0.61 g, 1.63 mmol) in ethanol (15 mL) wasadded ethyl 2-oxoacetate (50% solution in toluene, 0.37 mL, 1.80 mmol)and stirred at room temperature for 1 h. To this was added a solution ofNaCNBH₃ (0.12 g, 1.96 mmol) in ethanol (5 mL; containing 2 drops ofAcOH) dropwise over 10 min at room temperature and reaction mixture wasfurther stirred for 4 h. The reaction was monitored by TLC for completeconsumption of compound 8-i. Ethanol was removed under reduced pressure.The residue was treated with saturated NaHCO₃ solution (30 mL) andextracted with EtOAc (20 mL×3). The organic extract was washed withwater (20 mL×2) and brine (20 mL) then dried over anhydrous sodiumsulfate and concentrated under reduced pressure to afford 9-i (0.8 g,crude) which was carried forward to the next step without purification.LCMS: 459.20 (M+1).

Step 7: Synthesis of (R)—N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)-6-(2,4-dioxoimidazolidin-1-yl)hexane-1-sulfonamide (ProductionExample 26a)

To a stirred solution of compound 9-i (0.8 g, 1.74 mmol) in AcOH (15mL), was added KOCN (0.28 g, 3.49 mmol) and the reaction mixture wasstirred at room temperature for 16 h then heated at 60° C. for 6 h. Theprogress of the reaction was monitored by TLC. The reaction mixture wasconcentrated under reduced pressure and the residue treated with asaturated solution of NaHCO₃ solution (25 mL) and extracted with EtOAc(25 mL×2). The organic extract was washed with water (20 mL) and brine(20 mL) then dried over anhydrous sodium sulfate and concentrated underreduced pressure to dryness. The residue was purified by combiflashchromatography (eluting with 3-4% MeOH in DCM) to afford ProductionExample 26a as a white sticky solid (290 mg, 36.4% yield).

¹H NMR (400 MHz, DMSO-d₆) δ 10.67 (s, 1H), 7.62 (d, J=8.8 Hz, 1H),7.24-7.09 (m, 2H), 6.95-6.86 (m, 1H), 4.39 (p, J=7.2 Hz, 1H), 3.89 (d,J=4.5 Hz, 4H), 3.18-3.11 (m, 2H), 2.75-2.62 (m, 2H), 2.60-2.51 (m, 2H),1.55-1.32 (m, 8H), 1.32-1.02 (m, 2H), 0.59-0.55 (m, 2H), 0.34-0.25 (m,2H); ESI-MS (m/z): Calculated for: C₂₁H₃₀F₃O₅S: 455.55, observed mass;473.10 (M+10); HPLC purity: 99.39%.

Production Example 25a: Synthesis of(R)—N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)-4-(2,4-dioxoimidazolidin-1-yl)butane-1-sulfonamide

Step 1: Synthesis of 2-(4-bromobutyl)isoindoline-1,3-dione (3-j)

To a stirred solution of 1,4-dibromobutane (2-j) (9.7 mL, 27.0 mmol) inDMF (100 mL), was added potassium phthalate (1-j) (5.0 g, 27.0 mmol)portion-wise over 30 min at room temperature. After complete addition,the reaction mixture was stirred at 90° C. for 18 h, then quenched withwater (300 mL) and extracted with diethyl ether (150 mL×2). The combinedorganic extracts were washed with water (100 mL×2), followed by brine(50 mL×2) and dried over anhydrous sodium sulfate, filtered andconcentrated under reduced pressure to obtain the crude. The crudeproduct was purified by silica gel column chromatography (60-120 mesh)using 5-10% EtOAc/hexanes to afford 3-j as an off-white solid (5.5 g,72.3% yield). LCMS: 283.9 (M+1).

Step 2: Synthesis of S-(4-(1,3-dioxoisoindolin-2-yl)butyl) ethanethioate(4-j)

To a stirred solution of compound 3-j (5.5 g, 19.5 mmol) in DMF (50 mL),was added potassium thioacetate (2.67 g, 23.4 mmol) portion-wise over 20min at room temperature. After complete addition, the reaction mixturewas stirred at room temperature for 30 min. Reaction progress wasmonitored by TLC; after completion the reaction mixture was quenchedwith ice-cold water (250 mL) and stirred for 1 h. The precipitated wasfiltered, washed with water (100 mL) and dried in vacuo to afford 4-j asan off-white solid (5.0 g, 92.5%). LCMS: 278 (M+1).

Step 3: Synthesis of 4-(1,3-dioxoisoindolin-2-yl)butane-1-sulfonylchloride (5-j)

2N HCl (3.63 mL) was added to a stirred solution of compound 4-j (2.0 g,6.55 mmol) in acetonitrile (36 mL) at 0° C. This was followed byportion-wise addition of N-chlorosuccinimide (4.24 g, 31.7 mmol) over 30min and the reaction mixture was warmed to room temperature and stirredfor 1 h. Reaction progress was monitored by TLC. Upon completeconsumption of 4-j, reaction mixture was quenched with ice-cold water(100 mL) and extracted with diethyl ether (100 mL×2). The combinedorganic extracts were washed with saturated sodium bicarbonate solution(50 mL) and brine (50 mL) and dried over anhydrous sodium sulfate andconcentrated under reduced pressure to afford 5-j as an off-white solid(1.7 g, 78% yield).

¹H NMR (500 MHz, CDCl₃): δ 7.86-7.80 (m, 2H), 7.79-7.68 (m, 2H),3.76-3.65 (m, 4H), 2.09-2.0 (m, 2H), 1.92-1.80 (m, 2H).

Step 4: Synthesis of (R)—N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)-4-(1, 3-dioxoisoindolin-2-yl) butane-1-sulfonamide (7-j)

To a stirred solution of compound 6-j —HCl salt (0.5 g, 2.39 mmol) inCH₂Cl₂ (15 mL) was added triethylamine (0.9 mL, 6.33 mmol) at 0° C. andstirred. To this reaction mixture was added compound 5-j (1.08 mL, 3.58mmol) in CH₂Cl₂ (10 mL) dropwise over 25 min at 0° C. and stirred at thesame temperature for 3 h. The reaction was monitored by TLC for completeconsumption of compound 6-j. The reaction mixture was quenched withwater (50 mL) and extracted with CH₂Cl₂ (75 mL×2). The combined organicextracts were washed with brine (50 mL), dried over anhydrous sodiumsulfate and concentrated under reduced pressure. The crude was purifiedby silica gel flash chromatography using 15-20% EtOAc/hexanes to afford7-j as an off-white solid (1.0 g, 88.5%). LCMS: 492.10 (M+18).

Step 5: Synthesis of(R)-4-amino-N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl) ethyl)butane-1-sulfonamide (8-j)

To a stirred solution of compound 7-j (1.0 g, 2.10 mmol) in methanol (10mL) was added hydrazine hydrate (99%, 0.5 mL, 10.5 mmol) at 0° C. andstirred at room temperature for 3 h. The ice bath was removed and thereaction mixture was warmed to room temperature and stirred for 5 h. Thereaction was monitored by TLC till the complete consumption of compound7-j. After completion, methanol was removed from reaction mixture underreduced pressure. The crude material was dissolved in 2N HCl (50 mL) andwashed with diethyl ether (50 mL×3). The aqueous layer was basified withaq. ammonia (pH=˜8) and extracted with ethyl acetate (50 mL×2). Theorganic extracts were dried over anhydrous sodium sulfate andconcentrated under reduced pressure to afford 8-j as a sticky mass (0.64g, 88% yield). This product was carried forward to the next step withoutpurification. LCMS: 345.15 (M+1).

Step 6: Synthesis of ethyl(R)-(4-(N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl) ethyl) sulfamoyl)butyl)glycinate (9-j)

To a solution of compound 8-j (0.64 g, 1.86 mmol) in ethanol (15 mL) wasadded ethyl 2-oxoacetate (50% solution in toluene, 0.42 mL, 2.04 mmol)and stirred at room temperature for 1 h. To this was added a solution ofNaCNBH₃ (0.14 g, 2.23 mmol) in ethanol (5 mL; containing 2 drops ofAcOH) dropwise over 10 min then stirred at room temperature for 4 h. Thereaction was monitored by TLC for complete consumption of compound 8-j.Ethanol was removed under reduced pressure. The residue was taken insaturated NaHCO₃ solution (30 mL) and extracted with EtOAc (20 mL×3).The organic extract was washed with water (20 mL) and brine (20 mL)respectively then dried over anhydrous sodium sulfate and concentratedunder reduced pressure to afford 9-j (0.84 g, crude). It was carriedforward to the next step without purification. LCMS: 431.15 (M+1).

Step 7. Synthesis of (R)—N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)-4-(2,4-dioxoimidazolidin-1-yl)butane-1-sulfonamide (ProductionExample 25a)

To a stirred solution of compound 9-j (0.84 g, 1.95 mmol) in AcOH (15mL), was added KOCN (0.31 g, 3.90 mmol) and the reaction mixture wasstirred at room temperature for 16 h then heated at 60° C. for 6 h. Theprogress of the reaction was monitored by TLC. Reaction mixture wasconcentrated under reduced pressure and residue was taken in saturatedsolution of NaHCO₃ solution (20 mL) and extracted with EtOAc (20 mL×3).The organic extract was washed with water (25 mL×2) and brine (20 mL),dried over anhydrous sodium sulfate and concentrated under reducedpressure to dryness. The residue was purified by combiflashchromatography (eluting with 3-4% MeOH: DCM) to afford ProductionExample 25a as a white solid (130 mg, 15.5% yield).

¹H NMR (400 MHz, DMSO-d₆) δ 10.71 (s, 1H), 7.68 (d, J=8.7 Hz, 1H),7.22-7.09 (m, 2H), 6.91-6.85 (m, 1H), 4.45-4.33 (m, 1H), 3.93-3.83 (m,4H), 3.18-3.05 (m, 2H), 2.83-2.80 (m, 1H), 1.55-1.19 (m, 8H), 0.62-0.55(m, 2H), 0.38-0.30 (m, 2H); ESI-MS (m/z): Calculated for: C₁₉H₂₆FN₃O₅S:427.49, observed mass; 445.10 (M+18); HPLC purity: 99.7%.

Production Example 75a: Synthesis ofN-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-methylpropyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

Step 1: Synthesis of 4-fluoro-3-hydroxy-N-methoxy-N-methylbenzamide(2-k)

To a stirred solution of compound 1-k (10.0 g, 64.1 mmol) in DCM (270mL), was added triethylamine (8.1 mL, 57.6 mmol) followed by addition ofN,O-dimethylhydroxylamine.HCl (7.5 g, 76.9 mmol) and stirred at roomtemperature for 20 min. EDC.HCl (18.4 g, 96.1 mmol) was added at 0° C.and the reaction mixture was stirred at room temperature for 30 min(reaction deemed complete by TLC). The reaction mixture was quenchedwith NaHCO₃ solution and extracted with DCM. The combined organic layerwas washed with brine, dried over anhydrous sodium sulfate andconcentrated under reduced pressure to afford compound 2-k (10.0 g,crude). LCMS: 200.15 (M+1).

Step 2: Synthesis of3-(cyclopropylmethoxy)-4-fluoro-N-methoxy-N-methylbenzamide (4-k)

To a stirred solution of compound 2-k (10.0 g, 50.2 mmol) in ACN (100mL), was added CS₂CO₃ (24.5 g, 75.3 mmol) followed by addition of(bromomethyl)cyclopropane (7.21 mL, 75.3 mmol) and the reaction mixturewas refluxed for 5 h (reaction deemed complete by TLC). The reactionmixture was quenched with water and extracted with EtOAc. The combinedorganic layer was washed with brine, dried over anhydrous sodium sulfateand concentrated under reduced pressure. The residue was purified bycolumn chromatography using 25-30% EtOAc/hexane to afford compound 4-k(7.32 g, 57.6%). LCMS: 254 (M+1).

Step 3: Synthesis of1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-methylpropan-1-one (6-k)

To a stirred solution of compound 4-k (3.0 g, 11.8 mmol) in dry THF (50mL) was added isopropylmagnesium bromide (2.0 M in THF, 11.85 mL, 23.7mmol) at −10° C. The resultant mixture was stirred at room temperaturefor 12 h (reaction deemed complete by TLC). The reaction mixture wasquenched with NH₄Cl solution and diluted with EtOAc. The reactionmixture was filtered through a pad of Celite and extracted with EtOAc,washed with brine, dried over anhydrous sodium sulfate and evaporatedunder reduced pressure. The residue was purified by columnchromatography using 10% EtOAc/hexane to afford compound 6-k (1.1 g,39%). LCMS: 237.05 (M+1).

Step 4: Synthesis ofN-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-methylpropylidene)-2-methylpropane-2-sulfinamide(8-k)

To a stirred solution of compound 6-k (1.1 g, 4.66 mmol) and compound7-k (0.84 g, 6.99 mmol) in dry toluene (22 mL) was added Ti(O^(i)Pr)₄(2.65 g, 9.32 mmol). The resultant mixture was heated at reflux for 16 h(reaction deemed complete by TLC). The reaction mixture was filteredthrough a pad of Celite and the filtrate diluted with water andextracted with EtOAc. The combined organic layer was washed with brine,dried over anhydrous sodium sulfate and evaporated under reducedpressure. The residue was purified by column chromatography using 10%EtOAc/hexane to afford compound 8-k (0.82 g, 76%). LCMS: 340.10 (M+1).

Step 5: Synthesis ofN-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-methylpropyl)-2-methylpropane-2-sulfinamide(9-k)

To a stirred solution of DIBAL-H (1M solution in toluene, 6.1 mL, 6.04mmol) in dry toluene (10 mL), was added a solution of compound 8-k (0.82g, 2.41 mmol) in toluene (10 mL) dropwise at −78° C. The resultantmixture was stirred at −78° C. for 3 h (reaction deemed complete byTLC). The reaction mixture was quenched with NH₄Cl solution and dilutedwith EtOAc. The reaction mixture was filtered through a pad of Celiteand extracted with EtOAc, washed with brine, dried over anhydrous sodiumsulfate and evaporated under reduced pressure. The residue was purifiedby column chromatography using 10% EtOAc/hexane to afford compound 9-k(0.36 g, 60%). LCMS: 342.2 (M+1).

Step 6: Synthesis of3-(cyclopropylmethoxy)-4-fluorophenyl)-2-methylpropan-1-aminehydrochloride (10-k)

To a stirred solution of compound 9-k (0.67 g, 1.96 mmol) in MeOH (10mL), was added 4M HCl in dioxane (5 mL) and the resulting mixturestirred at room temperature for 3 h (reaction deemed complete by TLC).The reaction mixture was concentrated and the residue was purified bytrituration with diethyl ether to afford 10-k (0.44 g, 82%). LCMS: 274.1(M+1).

Step 7: Synthesis ofN-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-methylpropyl)-5-(1,3-dioxoisoindolin-2-yl)pentane-1-sulfonamide(12-k)

To a stirred solution of compound 10-k —HCl salt (0.42 g, 1.51 mmol) inDCM (10 mL) was added triethylamine (1.12 mL, 8.02 mmol) at 0° C. andstirred. To this reaction mixture was added compound 11-k (0.71 g, 2.27mmol) in DCM (5 mL) dropwise over 25 min at 0° C. and stirred at thesame temperature for 3 h. The reaction was monitored by TLC for completeconsumption of compound 11-k. The reaction mixture was quenched withwater (75 mL) and extracted with DCM (75 mL×2). The combined organicextracts were washed with brine (75 mL), dried over anhydrous sodiumsulfate and concentrated under reduced pressure. The crude was purifiedby silica gel flash chromatography using 15-20% EtOAc/hexanes to afford12-k as an off-white solid (0.69 g, 87%). LCMS: 517.25 (M+1).

Step 8: Synthesis of5-amino-N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-methyl propyl)pentane-1-sulfonamide (13-k)

To a stirred solution of compound 12-k (0.69 g, 1.33 mmol) in MeOH (7mL) was added hydrazine hydrate (99%, 0.33 mL, 6.68 mmol) at 0° C. Thenice bath was removed and the reaction mixture warmed to room temperatureand stirred for 3 h. The reaction was monitored by TLC till the completeconsumption of compound 12-k. After completion, MeOH was removed fromthe reaction mixture under reduced pressure. The resultant residue wasdissolved in 2N HCl (10 mL) and washed with diethyl ether (10 mL×3). Theaqueous layer was basified with aq. ammonia (pH=˜8) and extracted withEtOAc (10 mL×4). The organic extracts was dried over anhydrous sodiumsulfate and concentrated under reduced pressure to afford 13-k as asticky mass (0.5 g, crude). This material was carried forward to thenext step without purification. LCMS: 387.2 (M+1).

Step 9: Synthesis of ethyl(5-(N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-methylpropyl)sulfamoyl) pentyl) glycinate (15-k)

To a solution of compound 13-k (0.5 g, 1.29 mmol) in MeOH (15 mL), wasadded ethyl 2-oxoacetate (50% solution in toluene, 0.29 mL, 1.42 mmol)and stirred at room temperature for 1 h. To this was added a solution ofNaCNBH₃ (98 mg, 1.55 mmol) in ethanol (5 mL; containing 2 drops of AcOH)added dropwise over 10 min at room temperature and reaction mixture wasfurther stirred for 5 h. The reaction was monitored by TLC for completeconsumption of compound 13-k. Ethanol was removed under reducedpressure. The residue was treated with saturated NaHCO₃ solution (50 mL)and extracted with EtOAc (50 mL×2). The organic extract was washed withbrine (50 mL) then dried over anhydrous sodium sulfate and concentratedunder reduced pressure to afford 15-k (0.68 g, crude) which was carriedforward to the next step without purification. LCMS: 473.05 (M+1).

Step 10: Synthesis ofN-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-methylpropyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide(Production Example 75a)

To a stirred solution of compound 15-k (0.68 g, 1.44 mmol) in AcOH (12mL) was added KOCN (0.23 mg, 2.88 mmol) and the reaction mixture wasstirred at room temperature for 16 h then heated at 60° C. for 6 h. Theprogress of the reaction was monitored by TLC till complete consumptionof compound 15-k. The reaction mixture was concentrated under reducedpressure and treated with saturated solution of NaHCO₃ solution (50 mL)and extracted with EtOAc (50 mL×2). The organic extract was washed withwater (25 mL×2) and brine (20 mL) then dried over anhydrous sodiumsulfate and concentrated under reduced pressure to dryness. The residuewas purified by combiflash chromatography (eluting with 3-4% MeOH inDCM) to afford Production Example 75a as a white solid (10 mg, 2%yield).

¹H NMR (400 MHz, CDCl₃) δ 7.77 (s, 1H), 7.11-6.98 (m, 1H), 6.91-6.76 (m,2H), 5.13 (d, J=8.4 Hz, 1H), 4.03 (t, J=8.3 Hz, 1H), 4.00-3.81 (m, 4H),3.30-3.0 (m, 2H), 2.69-2.60 (m, 1H), 2.52-2.43 (m, 1H), 1.95-1.80 (m,1H), 1.40-1.29 (m, 4H), 1.27 (d, J=15.0 Hz, 3H), 1.19 (d, J=11.3 Hz,1H), 1.06 (d, J=6.6 Hz, 3H), 0.80 (d, J=6.5 Hz, 2H), 0.66 (d, J=8.0 Hz,2H), 0.36 (d, J=8.1 Hz, 2H); ESI-MS (m/z): Calculated for: C₂₂H₃₂FN₃O₅S:469.57, observed mass; 470.25 (M+H); HPLC purity: 97.22%.

Step 11: Synthesis of 2-(5-bromopentyl) isoindoline-1,3-dione (14-k)

To a stirred solution of 1,5-dibromopentane (13-k) (170.58 mL, 1.26 mol)in DMF (1.5 L) was added potassium phthalate (12-k) (78.0 g, 0.42 mol)portion-wise over 30 min at room temperature. After complete addition,the reaction mixture was stirred at 90° C. for 18 h, then quenched withwater (3 L) and extracted with diethyl ether (500 mL×4). The combinedorganic extracts were washed with water (500 mL×2), followed by brine(500 mL×2) and dried over anhydrous sodium sulfate, filtered andconcentrated under reduced pressure to obtain the crude. The residue waspurified by silica gel column chromatography (60-120 mesh) using 5-10%EtOAc/hexanes to afford 14-k as an off-white solid (81 g, 65% yield).

¹H NMR (400 MHz, CDCl₃): δ 7.82 (dd, J=5.5, 3.1 Hz, 2H), 7.69 (dd,J=5.5, 3.0 Hz, 2H), 3.68 (t, J=7.2 Hz, 2H), 3.38 (t, J=6.8 Hz, 2H),1.93-1.85 (m, 2H), 1.70 (p, J=7.5 Hz, 2H), 1.53-1.43 (m, 2H).

Step 12: Synthesis ofS-(5-(1,3-dioxoisoindolin-2-yl)pentyl)ethanethioate (15-k)

To a stirred solution of compound 14-k (80 g, 0.27 mol) in DMF (745 mL)was added potassium thioacetate (34 g, 0.29 mol) portion wise over 20min at room temperature. After complete addition, the reaction mixturewas stirred at room temperature for 30 min. Reaction progress wasmonitored by TLC; after completion the reaction mixture was quenchedwith ice-cold water (1 L) and stirred for 1 h. The resultant precipitatewas filtered, washed with water (500 mL) and dried in vacuo to afford15-k as an off-white solid (75 g, 95%).

¹H NMR (400 MHz, CDCl₃): δ 7.84 (dd, J=5.5 Hz, 3.0 Hz, 2H), 7.71 (dd,J=5.5 Hz, 3.1 Hz, 2H), 3.67 (t, J=7.3 Hz, 2H), 2.85 (t, J=7.3 Hz, 2H),2.30 (s, 3H), 1.73-1.65 (m, 3H), 1.64-1.57 (m, 1H), 1.46-1.37 (m, 2H);LC-MS: 292.2 (M⁺+1).

Step 13: Synthesis of 5-(1,3-dioxoisoindolin-2-yl)pentane-1-sulfonylchloride (11-k)

2N HCl (135 mL) was added to a stirred solution of compound 15-k (74 g,0.25 mol) in acetonitrile (1.35 L) at 0° C. This was followed byportion-wise addition of N-chlorosuccinimide (135.8 g, 1.10 mol) over 30min and the reaction mixture was warmed to room temperature and stirredfor 1 h. Reaction progress was monitored by TLC. Upon completeconsumption of 15-k, reaction mixture was quenched with ice-cold water(500 mL) and extracted with diethyl ether (500 mL×2). The combinedorganic extracts were washed with saturated sodium bicarbonate solution(500 mL), water (500 mL) and brine (500 mL) and dried over anhydroussodium sulfate and concentrated under reduced pressure to afford 11-k asan off-white solid (73 g, 91% yield).

¹H NMR (500 MHz, CDCl₃): δ 7.85 (dd, J=5.5 Hz, 3.2 Hz, 2H), 7.72 (dd,J=3.2 Hz, 5.5 Hz, 2H), 3.72 (t, J=7.0 Hz, 2H), 3.68-3.63 (m, 2H),2.15-2.05 (m, 2H), 1.77 (m, 2H), 1.62-1.52 (m, 2H); LC-MS: 316.1 (M⁺+1).

Production Example 76a: Synthesis ofN-(1-(3-(2-cyclopropylethoxy)-4-fluorophenyl) ethyl)-5-(2,4-dioxoimidazolidin-1-yl) pentane-1-sulfonamide

Step 1: Synthesis of 1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethan-1-one (3-l)

To a stirred solution of 1-l (5.0 g, 32.4 mmol) in DMF (50 mL) was addedK₂CO₃ (11.2 g, 81.1 mmol) followed by addition of(bromomethyl)cyclopropane (3.73 g, 38.9 mmol) and the reaction mixturewas refluxed for 2 h (reaction deemed complete by TLC). The reactionmixture was quenched with water and extracted with EtOAc. The combinedorganic layer was washed with brine, dried over anhydrous sodium sulfateand concentrated under reduced pressure to afford 3-l (6.61 g, crude).LCMS: 208.95 (M+1).

Step 2: Synthesis of1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2,2-dimethylpropan-1-one (5-l)

To a stirred solution of compound 3-l (2.2 g, 9.36 mmol) in dry toluene(30 mL), KOH (5.24 g, 93.6 mmol) was added and stirred at roomtemperature for 10 min. 18-crown-6 ether (1.23 g, 4.68 mmol) was addedfollowed by addition of methyl iodide (26.5 g, 187 mmol) and theresultant mixture heated at 70° C. for 48 h (reaction deemed complete byTLC). The reaction mixture was cooled to room temperature and dilutedwith ice-water and extracted with EtOAc. The combined organic layer waswashed with brine, dried over anhydrous sodium sulfate and evaporatedunder reduced pressure. The residue was purified by columnchromatography to afford a mixture of compound 5-l and1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-methylpropan-1-one (1.0 g,42.7%). LCMS: 251.2 (M+1).

Step 3: Synthesis ofN-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2,2-dimethylpropylidene)-2-methylpropane-2-sulfinamide(7-l)

To a stirred solution of compound 5-l (1.0 g, 4.0 mmol) and compound 6-l(0.77 g, 6.4 mmol) in dry toluene (5 mL) was added Ti(O^(i)Pr)₄ (3.40 g,12.0 mmol). The resultant mixture was heated at 100° C. for 16 h(reaction deemed complete by TLC). The reaction mixture was diluted withEtOAc and quenched with water and filtered through a pad of Celite. Thefiltrate was diluted with water and extracted with EtOAc. The combinedorganic layer was washed with brine, dried over anhydrous sodium sulfateand evaporated under reduced pressure. The residue was purified bycolumn chromatograph to afford mixture of compound 7-l andN-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-methylpropylidene)-2-methylpropane-2-sulfinamide(1.0 g, crude). LCMS: 354 (M+1).

Step 4: Synthesis ofN-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2,2-dimethylpropyl)-2-methylpropane-2-sulfinamide(8-l)

To a stirred solution of DIBAL-H (1M solution in toluene, 8.0 mL, 8.84mmol) in dry toluene (5 mL), was added a solution of compound 7-l (1.0g, 2.83 mmol) in toluene (5 mL) dropwise at −78° C. The resultantmixture was stirred at same temperature for 3 h (reaction deemedcomplete by TLC). The reaction mixture was quenched with NH₄Cl solutionand diluted with EtOAc. The reaction mixture was filtered through a padof Celite and extracted with EtOAc, washed with brine, dried overanhydrous sodium sulfate and evaporated under reduced pressure to affordcompound8-1N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2-methylpropyl)-2-methylpropane-2-sulfinamide(1.0 g, crude). LCMS: 342.05 (M+1).

Step 5: Synthesis of1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2,2-dimethylpropan-1-aminehydrochloride (9-l)

To a stirred solution of compound 8-l (1.0 g, 2.81 mmol) in MeOH (5 mL),was added 4M HCl in dioxane (1 mL) and the resulting mixture stirred atroom temperature for 3 h (reaction deemed complete by TLC). The reactionmixture was concentrated and the residue was purified by triturationwith methyl tert butyl ether to afford mixture of compound 9-1 (0.5 g,crude) as a colorless solid. LCMS: 236.1 (M+1).

Step 6: Synthesis ofN-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2,2-dimethylpropyl)-5-(1,3-dioxoisoindolin-2-yl)pentane-1-sulfonamide(11-1)

To a stirred solution of compound 9-l —HCl salt (0.5 g, 2.12 mmol) inDCM (5 mL) was added triethylamine (1.0 mL, 10.6 mmol) at 0° C. andstirred. To this reaction mixture was added compound 10-1 (1.0 g, 3.19mmol) in DCM (5 mL) dropwise over 25 min at 0° C. and stirred at thesame temperature for 3 h. The reaction was monitored by TLC for completeconsumption of compound 9-l. The reaction mixture was quenched withwater (50 mL) and extracted with DCM (50 mL×2). The combined organicextracts were washed with brine (50 mL), dried over anhydrous sodiumsulfate and concentrated under reduced pressure to afford compound 11-las an off-white solid (0.4 g, crude). LCMS: 531.1 (M+1).

Step 7: Synthesis of5-amino-N-(1-(3-(2-cyclopropylethoxy)-4-fluorophenyl)ethyl)pentane-1-sulfonamide(12-l)

To a stirred solution of compound 11-l (0.4 g, 0.75 mmol) in ethanol (5mL) was added hydrazine hydrate (99%, 0.18 mL, 3.77 mmol) at 0° C. andstirred at room temperature for 3 h. The ice bath was removed and thereaction mixture warmed to room temperature and stirred for 5 h. Thereaction was monitored by TLC till the complete consumption of compound11-l. After completion, ethanol was removed from reaction mixture underreduced pressure. The crude material was dissolved in 2N HCl (25 mL) andwashed with diethyl ether (25 mL×2). The aqueous layer was basified withsaturated aqueous ammonia (pH=˜8) and extracted with EtOAc (25 mL×2).The organic extracts were dried over anhydrous sodium sulfate andconcentrated under reduced pressure to afford mixture of compound 12-las a yellow sticky solid (0.2 g, crude). This material was carriedforward to the next step without purification. LCMS: 401 (M+1).

Step 8: Synthesis of ethyl(5-(N-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)-2,2-dimethylpropyl)sulfamoyl)pentyl)glycinate(14-l)

To a solution of compound 12-l (0.2 g, 0.5 mmol) in ethanol (8 mL) wasadded ethyl 2-oxoacetate (50% solution in toluene, 56 mg, 0.55 mmol) andstirred at room temperature for 1 h. To this was added a solution ofNaCNBH₃ (37.2 mg, 0.6 mmol) in ethanol (8 mL; containing 2 drops ofAcOH) added dropwise over 10 min to the reaction mixture at roomtemperature and reaction mixture was further stirred for 4 h. Thereaction was monitored by TLC for complete consumption of compound 11-l.Ethanol was removed under reduced pressure. The residue was treated withsaturated NaHCO₃ solution (15 mL) and extracted with EtOAc (20 mL×2).The organic extract was washed with water (20 mL) and brine (15 mL)respectively then dried over anhydrous sodium sulfate and concentratedunder reduced pressure to afford compound 14-l (0.15 g, crude), whichwas carried forward to the next step without purification. LCMS: 487.1(M+1).

Step 9: Synthesis of N-(1-(3-(2-cyclopropylethoxy)-4-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl) pentane-1-sulfonamide (ProductionExample 76a)

To a stirred solution of compound 14-l (0.15 g, 0.30 mmol) in AcOH (5mL) was added KOCN (50 mg, 0.61 mmol) and the reaction mixture wasstirred at room temperature for 16 h then heated at 60° C. for 6 h. Theprogress of the reaction was monitored by TLC till complete consumptionof compound 14-l. The reaction mixture was concentrated under reducedpressure and the residue treated with saturated aqueous NaHCO₃ (50 mL)and extracted with EtOAc (50 mL×2). The organic extract was washed withwater (50 mL) and brine (15 mL) respectively then dried over anhydroussodium sulfate and concentrated under reduced pressure to dryness. Theresidue was purified by flash column chromatography (eluting with 3-4%MeOH in DCM) to afford Production Example 76a as a white solid (20 mg,14%).

¹H NMR (400 MHz, DMSO-d₆) δ 10.68 (s, 1H), 7.65 (d, J=10.7 Hz, 1H), 7.25(d, J=8.6 Hz, 1H), 7.12 (dd, J=11.4, 8.2 Hz, 1H), 6.91 (s, 1H), 5.76 (d,J=1.8 Hz, 1H), 4.00 (d, J=10.6 Hz, 1H), 3.92-3.85 (m, 2H), 3.84-3.81 (m,2H), 3.10-3.01 (m, 2H), 2.65 (d, J=4.2 Hz, 1H), 2.35 (d, J=4.2 Hz, 1H),2.00 (q, J=7.1 Hz, 1H), 1.34 (d, J=1.7 Hz, 3H), 1.06-0.99 (m, 1H), 0.86(s, 9H), 0.58-0.55 (m, 2H), 0.36-0.28 (m, 2H); ESI-MS (m/z): Calculatedfor: C₂₃H₃₄FN₃O₅S: 483.60, observed mass; LCMS purity: 482.15 (M−H).

Production Example 77a: Synthesis of N-(cyclopropyl(3-(cyclopropylmethoxy)-4-fluorophenyl) methyl)-5-(2,4-dioxoimidazolidin-1-yl) pentane-1-sulfonamide

Step 1: Synthesis of 4-fluoro-3-hydroxy-N-methoxy-N-methylbenzamide(2-m)

To a stirred solution of compound 1-m (10.0 g, 64.1 mmol) in DCM (120mL), was added triethylamine (17 g, 121 mmol) followed by addition ofN,O-dimethylhydroxylamine.HCl (7.5 g, 76.9 mmol) and stirred at roomtemperature for 20 min. EDC.HCl (18.4 g, 96.1 mmol) was added at 0° C.and the reaction mixture was stirred at room temperature for 12 h(reaction deemed complete by TLC). The reaction mixture was quenchedwith NaHCO₃ solution and extracted with DCM. The combined organic layerwas washed with brine, dried over anhydrous sodium sulfate andconcentrated under reduced pressure to afford compound 2-m (10 g,crude). LCMS: 200.15 (M+1).

Step 2: Synthesis of3-(cyclopropylmethoxy)-4-fluoro-N-methoxy-N-methylbenzamide (4-m)

To a stirred solution of compound 2-m (10.0 g, 50.2 mmol) in DMF (100mL) was added K₂CO₃ (13.8 g, 100 mmol) followed by addition of(bromomethyl)cyclopropane (8.14 g, 60.3 mmol) and the reaction mixturewas refluxed for 16 h (reaction deemed complete by TLC). The reactionmixture was quenched with water and extracted with EtOAc. The combinedorganic layer was washed with brine, dried over anhydrous sodium sulfateand concentrated under reduced pressure. The residue was purified bycolumn chromatography using 25-30% EtOAc/hexane to afford compound 4-m(10 g, 78.6%). LCMS: 253.9 (M+1).

Step 3: Synthesis of 1-(3-(cyclopropylmethoxy)-5-fluorophenyl)ethan-1-one (6-m)

To a stirred solution of compound 4-m (4.0 g, 15.8 mmol) in dry THF (30mL) was added freshly prepared cyclopropylmagnesium bromide (20.5 mL,20.5 mmol) at −10° C. The resultant mixture was stirred at roomtemperature for 12 h (reaction deemed complete by TLC). The reactionmixture was quenched with saturated NH₄Cl solution and diluted withEtOAc. The reaction mixture was filtered through a pad of Celite andextracted with EtOAc, washed with brine, dried over anhydrous sodiumsulfate and evaporated under reduced pressure. The residue was purifiedby column chromatography using 15% EtOAc/hexane to afford compound 6-m(1.5 g, 40.5%). LCMS: 234.95 (M+1).

Step 4: Synthesis ofN-(cyclopropyl(3-(cyclopropylmethoxy)-4-fluorophenyl)methylene)-2-methylpropane-2-sulfinamide(8-m)

To a stirred solution of compound 6-m (1.5 g, 6.41 mmol) and compound7-m (1.24 g, 10.2 mmol) in dry toluene (25 mL) was added Ti(O^(i)Pr)₄(3.79 mL, 12.8 mmol). The resultant mixture was refluxed for 16 h(reaction deemed complete by TLC). The reaction mixture was diluted withEtOAc and quenched with water and filtered through a pad of Celite. Thefiltrate was diluted with water and extracted with EtOAc. The combinedorganic layer was washed with brine, dried over anhydrous sodium sulfateand evaporated under reduced pressure. The residue was purified bycolumn chromatography using 10% EtOAc/hexane to afford compound 8-m (1.2g, crude). LCMS: 338.05 (M+1).

Step 5: Synthesis of N-(cyclopropyl(3-(cyclopropylmethoxy)-4-fluorophenyl)methyl)-2-methylpropane-2-sulfinamide (9-m)

To a stirred solution of DIBAL-H (1M solution in toluene, 1.78 mL, 1.78mmol) in dry toluene (3 mL), was added a solution of compound 8-m (0.3g, 0.89 mmol) in toluene (3 mL) dropwise at −78° C. The resultantmixture was stirred at −78° C. for 3 h (reaction deemed complete byTLC). The reaction mixture was quenched with saturated NH₄Cl solutionand extracted with EtOAc. The organic layer was separated and washedwith brine, dried over anhydrous sodium sulfate and evaporated underreduced pressure. The residue was purified by column chromatographyusing 10% EtOAc/hexane to afford compound 9-m (0.1 g, 33%). LCMS: 340.05(M+1).

Step 6: Synthesis of (R)-cyclopropyl(3-(cyclopropylmethoxy)-4-fluorophenyl) methanamine hydrochloride (10-m)

To a stirred solution of compound 9-m (0.1 g, 0.29 mmol) in dioxane (5mL), was added 4M HCl in dioxane (5 mL) and the resulting mixturestirred at room temperature for 3 h (reaction deemed complete by TLC).The reaction mixture was concentrated and the residue was purified bytrituration with diethyl ether to afford compound 10-m (75 mg, crude).LCMS: 237.1 (M+1).

Step 7: Synthesis of (R)—N-(cyclopropyl(3-(cyclopropylmethoxy)-4-fluorophenyl)methyl)-5-(1,3-dioxoisoindolin-2-yl)pentane-1-sulfonamide (12-m)

To a stirred solution of compound 10-m —HCl salt (0.075 g, 0.27 mmol) inDCM (2 mL) was added triethylamine (0.19 mL, 1.38 mmol) at 0° C. andstirred. To this reaction mixture was added compound 11-m (0.13 g, 0.41mmol) in DCM (2 mL) dropwise over 25 min at 0° C. and stirred at roomtemperature for 3 h. The reaction was monitored by TLC for completeconsumption of compound 10-m. The reaction mixture was quenched withwater (25 mL) and extracted with DCM (25 mL×2). The combined organicextracts was washed with brine (25 mL), dried over anhydrous sodiumsulfate and concentrated under reduced pressure. The crude was purifiedby combiflash chromatography using 15-20% EtOAc/hexanes to affordcompound 12-m (0.1 g, 70.4%). LCMS: 513.15 (M-1).

Step 8: Synthesis of 5-amino-N-(cyclopropyl(3-(cyclopropylmethoxy)-4-fluorophenyl) methyl) pentane-1-sulfonamide(13-m)

To a stirred solution of compound 12-m (0.1 g, 0.19 mmol) in MeOH (1mL), was added hydrazine hydrate (99%, 48 μL, 0.97 mmol) at 0° C. andthen ice bath was removed. The reaction mixture was warmed to roomtemperature and stirred for 3 h. The reaction was monitored by TLC tillthe complete consumption of compound 12-m. After completion, MeOH wasremoved from the reaction mixture under reduced pressure. The residuewas dissolved in 2N HCl (10 mL) and washed with diethyl ether (10 mL×3).The aqueous layer was basified with saturated aqueous ammonia (pH=˜8)and extracted with EtOAc (10 mL×4). The organic extracts was dried overanhydrous sodium sulfate and concentrated under reduced pressure toafford compound 13-m (70 mg, crude). This product was carried forward tothe next step without purification. LCMS: 385.1 (M+1).

Step 9: Synthesis of ethyl (5-(N-(cyclopropyl(3-(cyclopropylmethoxy)-4-fluorophenyl) methyl) sulfamoyl) pentyl)glycinate (15-m)

To a solution of compound 13-m (0.07 g, 0.18 mmol) in ethanol (3 mL) wasadded compound 14-m (50% solution in toluene, 20 μL, 0.2 mmol) andstirred at room temperature for 1 h. To this was added a solution ofNaCNBH₃ (13 mg, 0.21 mmol) in ethanol (2 mL containing 1 drop of AcOH)dropwise over 10 min to the reaction mixture at room temperature andreaction mixture was further stirred for 5 h. The reaction was monitoredby TLC for complete consumption of compound 13-m. Ethanol was removedunder reduced pressure. The residue was taken in saturated NaHCO₃solution (25 mL) and extracted with EtOAc (25 mL×2). The organic extractwas washed with brine (25 mL) respectively then dried over anhydroussodium sulfate and concentrated under reduced pressure to affordcompound 15-m (70 mg, crude) which was carried forward to the next stepwithout purification. LCMS: 471.15 (M+1).

Step 10: Synthesis of N-(cyclopropyl(3-(cyclopropylmethoxy)-4-fluorophenyl) methyl)-5-(2,4-dioxoimidazolidin-1-yl) pentane-1-sulfonamide (Production Example 77a)

To a stirred solution of compound 15-m (0.07 g, 0.14 mmol) in AcOH (2mL) was added KOCN (24 mg, 0.29 mmol) and the reaction mixture wasstirred at room temperature for 16 h. The reaction mixture was heated at60° C. for 6 h. The progress of the reaction was monitored by TLC.Reaction mixture was concentrated under reduced pressure and residue wastaken in saturated solution of NaHCO₃ (25 mL) and extracted with EtOAc(25 mL×2). The organic extract was washed with brine (10 mL), then driedover anhydrous sodium sulfate, filtered and concentrated under reducedpressure to dryness. The residue was purified by Prep-HPLC to affordProduction Example 77a as a sticky solid (5 mg, HPLC purity: 77%).

Production Example 78a: Synthesis of((S)—N-(1-(3-(cyclopropylmethoxy)-5-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide

Step 1: Synthesis of 3-fluoro-5-hydroxy-N-methoxy-N-methylbenzamide(2-n)

To a stirred solution of compound 1-n (3.0 g, 19.2 mmol) in DCM (30 mL),was added triethylamine (5.5 mL, 28.0 mmol) followed by addition ofN,O-dimethylhydroxylamine.HCl (2.24 g, 23.0 mmol) and stirred at roomtemperature for 20 min. EDC.HCl (5.53 g, 28.0 mmol) was added at 0° C.and the reaction mixture was stirred at room temperature for 30 min(reaction deemed complete by TLC). The reaction mixture was quenchedwith NaHCO₃ solution and extracted with DCM. The combined organic layerwas washed with brine, dried over anhydrous Na₂SO₄ and concentratedunder reduced pressure to afford compound 2-n (3.8 g, crude). LCMS: 200(M+1).

Step 2: Synthesis of3-(cyclopropylmethoxy)-5-fluoro-N-methoxy-N-methylbenzamide (4-n)

To a stirred solution of compound 2-n (3.8 g, 19.2 mmol) in ACN (30 mL),was added Cs₂CO₃ (9.1 g, 28.0 mmol) followed by addition of(bromomethyl)cyclopropane (3-n) (3.8 g, 28.0 mmol) and the reactionmixture was refluxed for 5 h (reaction deemed complete by TLC). Thereaction mixture was quenched with water and extracted with EtOAc. Thecombined organic layer was washed with brine, dried over anhydrousNa₂SO₄ and concentrated under reduced pressure. The residue was purifiedby column chromatography using 25-30% EtOAc/hexane to afford compound4-n (2.6 g, 54.1%). LCMS: 254 (M+1).

Step 3: Synthesis of1-(3-(cyclopropylmethoxy)-5-fluorophenyl)ethan-1-one (6-n)

To a stirred solution of compound 4-n (2.6 g, 10.2 mmol) in dry THF (25mL) was added methyl magnesium bromide (5-n) (3.0 M in THF, 6.84 mL,20.5 mmol) at −10° C. The resultant mixture was stirred at sametemperature for 12 h (reaction deemed complete by TLC). The reactionmixture was quenched with NH₄Cl solution and diluted with EtOAc. Thereaction mixture was filtered through a pad of Celite and extracted withEtOAc, washed with brine, dried over anhydrous Na₂SO₄ and evaporatedunder reduced pressure. The residue was purified by columnchromatography using 10% EtOAc/hexane to afford compound 6-n (1.8 g,84%). LCMS: 209 (M+1).

Step 4: Synthesis of(S)—N-(1-(3-(cyclopropylmethoxy)-5-fluorophenyl)ethylidene)-2-methylpropane-2-sulfinamide (8-n)

To a stirred solution of compound 6-n (1.8 g, 8.65 mmol) and compound7-n (1.78 g, 14.7 mmol) in dry toluene (20 mL) was added Ti(O^(i)Pr)₄(6.83 mL, 21.6 mmol). The resultant mixture was heated at 90° C. for 16h (reaction deemed complete by TLC). The reaction mixture was filteredthrough a pad of Celite, the filtrate was diluted with water andextracted with EtOAc. The combined organic layer was washed with brine,dried over anhydrous Na₂SO₄ and evaporated under reduced pressure. Theresidue was purified by column chromatography using 10% EtOAc/hexane toafford compound 8-n (2.6 g, crude). LCMS: 312.15 (M+1).

Step 5: Synthesis of(S)—N—((S)-1-(3-(cyclopropylmethoxy)-5-fluorophenyl) ethyl)-2-methylpropane-2-sulfinamide (9-n)

To a stirred solution of DIBAL-H (1M solution in toluene, 41.7 mL, 41.75mmol) in dry toluene (20 mL), was added a solution of compound 8-n (2.6g, 8.35 mmol) in toluene (10 mL) dropwise at −78° C. The resultantmixture was stirred at same temperature for 3 h (reaction deemedcomplete by TLC). The reaction mixture was quenched with NH₄Cl solutionand diluted with EtOAc. The reaction mixture was filtered through a padof Celite and extracted with EtOAc, washed with brine, dried overanhydrous Na₂SO₄ and evaporated under reduced pressure. The residue waspurified by column chromatography using 10% EtOAc/hexane to affordcompound 9-n (1.2 g, 45.8%). LCMS: 314.05 (M+1).

Step 6: Synthesis of (S)-1-(3-(cyclopropylmethoxy)-5-fluorophenyl)ethan-1-amine hydrochloride (10-n)

To a stirred solution of compound 9-n (1.2 g, 3.83 mmol) in dioxane (5mL), was added 4M HCl in dioxane (10 mL) and the resulting mixturestirred at room temperature for 3 h (reaction deemed complete by TLC).The reaction mixture was concentrated and the residue was purified bytrituration with diethyl ether to afford 10-n (1.3 g, crude). LCMS: 210(M+1).

Step 7: Synthesis of (S)—N-(1-(3-(cyclopropylmethoxy)-5-fluorophenyl)ethyl)-5-(1,3-dioxoisoindolin-2-yl) pentane-1-sulfonamide (12-n)

To a stirred solution of compound 10-n —HCl salt (1.3 g, 5.29 mmol) inCH₂Cl₂ (10 mL) was added triethylamine (2.21 mL, 15.8 mmol) at 0° C. andstirred. To this reaction mixture was added compound 11-n (2.51 g, 7.94mmol) in CH₂Cl₂ (10 mL) dropwise over 25 min at 0° C. and stirred at thesame temperature for 3 h. The reaction was monitored by TLC for completeconsumption of compound 10-n. The reaction mixture was quenched withwater (100 mL) and extracted with CH₂Cl₂ (100 mL×2). The combinedorganic extracts was washed with brine (100 mL), dried over anhydroussodium sulfate and concentrated under reduced pressure. The crude waspurified by silica gel flash chromatography using 15-20% EtOAc/hexanesto afford 12-n as an off-white solid (1.5 g, 58.1%). LCMS: 489.1 (M+1).

Step 8: Synthesis of(S)-5-amino-N-(1-(3-(cyclopropylmethoxy)-5-fluorophenyl) ethyl)pentane-1-sulfonamide (13-n)

To a stirred solution of compound 12-n (1.5 g, 3.07 mmol) in methanol(20 mL), was added hydrazine hydrate (99%, 0.48 mL, 15.3 mmol) at 0° C.and stirred at room temperature for 3 h. The ice bath was removed andthe reaction mixture was warmed to room temperature and stirred for 5 h.The reaction was monitored by TLC till the complete consumption ofcompound 12-n. After completion, methanol was removed from reactionmixture under reduced pressure. The crude material was dissolved in 2NHCl (10 mL) and washed with diethyl ether (10 mL×3). The aqueous layerwas basified with aq. ammonia (pH=˜8) and extracted with ethyl acetate(10 mL×4). The organic extracts was dried over anhydrous sodium sulfateand concentrated under reduced pressure to afford 13-n as a sticky mass(1.1 g, crude). This product was carried forward to the next stepwithout purification. LCMS: 359 (M+1).

Step 9: Synthesis of ethyl(S)-(5-(N-(1-(3-(cyclopropylmethoxy)-5-fluorophenyl) ethyl) sulfamoyl)pentyl) glycinate (15-n)

To a solution of compound 13-n (0.6 g, 1.68 mmol) in ethanol (10 mL) wasadded ethyl 2-oxoacetate (14-n) (50% solution in toluene, 0.36 mL, 1.85mmol) and stirred at room temperature for 1 h. To this was added asolution of NaCNBH₃ (426 mg, 2.02 mmol) in ethanol (5 mL; containing 2drops of AcOH) dropwise over 10 min at room temperature and reactionmixture was stirred for another 4 h. The reaction was monitored by TLCfor complete consumption of compound 13-n. Ethanol was removed underreduced pressure. The residue was taken in saturated NaHCO₃ solution (75mL) and extracted with EtOAc (75 mL×2). The organic extract was washedwith brine (75 mL) respectively then dried over anhydrous sodium sulfateand concentrated under reduced pressure to afford 15-n (0.8 g, crude).It was carried forward to the next step without purification.

Step 10: Synthesis of ((S)—N-(1-(3-(cyclopropylmethoxy)-5-fluorophenyl)ethyl)-5-(2,4-dioxoimidazolidin-1-yl)pentane-1-sulfonamide (ProductionExample 78a)

To a stirred solution of compound 15-n (0.8 g, 1.86 mmol) in AcOH (10mL) was added KOCN (0.3 mg, 3.72 mmol) and the reaction mixture wasstirred at room temperature for 16 h then heated at 60° C. for 6 h. Theprogress of the reaction was monitored by TLC. The reaction mixture wasconcentrated under reduced pressure and residue was taken in saturatedsolution of NaHCO₃ solution (100 mL) and extracted with EtOAc (75 mL×2).The organic extract was washed with brine (20 mL) then dried overanhydrous sodium sulfate and concentrated under reduced pressure todryness. The residue was purified by combiflash chromatography (elutingwith 3-4% MeOH in DCM) to afford Production Example 78a as a white solid(0.22 g, 26%). ¹H NMR (400 MHz, DMSO-d₆) δ 10.68 (s, 1H), 7.69 (d, J=8.8Hz, 1H), 6.83-6.72 (m, 2H), 6.64-6.60 (m, 1H), 4.45-4.33 (m, 1H), 3.86(s, 2H), 3.80 (d, J=7.0 Hz, 2H), 3.13 (t, J=7.0 Hz, 2H), 2.82-2.78 (m,1H), 2.60-2.55 (m, 1H), 1.61-1.41 (m, 2H), 1.34 (t, J=6.3 Hz, 4H),1.26-1.02 (m, 4H), 0.60-0.49 (m, 2H), 0.34-0.22 (m, 2H); ESI-MS (m/z):Calculated for: C₂₀H₂₈FN₃O₅S: 441.52, observed mass; 440.1 (M−H); HPLCpurity: 99.1%.

Example 79: Synthesis of(5)-1-(4-(2-(hydroxydiphenylmethyl)pyrrolidin-1-yl)-4-oxobutyl)imidazolidine-2,4-dione(alternate ID: 1229)

Step-1: Synthesis of 1-(tert-butyl) 2-methyl(S)-pyrrolidine-1,2-dicarboxylate (2)

To a stirred solution of 1 (0.8 g, 3.71 mmol) in dry THF (25 mL), K₂CO₃(0.77 g, 5.57 mmol) was added at 0° C. and stirred at the sametemperature for 10 min. To the reaction mixture methyl iodide (0.79 g,5.57 mmol) was added and the reaction mixture was stirred at roomtemperature for 16 h. The progress of the reaction was monitored by TLCfor complete consumption of compound 1. All volatiles were removed underreduced pressure to afford crude. The crude product was purified bysilica gel column chromatography (100-200 mesh) using 12% EtOAc/hexanesto afford 2 as off-white solid (0.4 g, 47% yield). LCMS: 129.90 (M-Boc).¹H NMR (400 MHz, DMSO-d₆) δ 1.32 (s, 9H), 1.76-1.87 (m, 2H), 2.15-2.26(m, 2H), 3.33-3.37 (m, 2H), 3.65 (s, 3H), 4.13-4.17 (m, 1H).

Step-2: Synthesis of tert-butyl(S)-2-(hydroxydiphenylmethyl)pyrrolidine-1-carboxylate (3)

A stirred solution of compound (2) (0.38 g, 1.65 mmol) in THF (25 mL) at0° C. was treated with phenyl magnesium bromide (3M solution in diethylether (1.8 mL, 26.2 mmol) and the reaction mixture allowed to warm toroom temperature and stirred for a further 16 h. Progress of thereaction was monitored by TLC. The reaction mixture was quenched withsaturated NH₄Cl solution and extracted with EtOAc (10 mL×2). Combinedorganic layer was washed with water, brine, dried over sodium sulphateand concentrated under reduced pressure. The crude product was purifiedby silica gel column chromatography (100-200 mesh) using 10% EtOAc inhexanes to afford 3 as an off-white solid (0.35 g, 60% yield). LCMS:352.75 (M−H). ¹H NMR (400 MHz, DMSO-d₆) δ 1.08 (s, 9H), 1.60-1.62 (m,1H), 1.70-1.79 (m, 1H), 1.83-1.97 (m, 2H), 3.58 (brs, 1H), 4.83 (d,J=6.85 Hz, 1H), 5.67 (s, 1H), 7.11-7.16 (m, 1H), 7.19 (t, J=7.34 Hz,2H), 7.24-7.30 (m, 3H), 7.35 (t, J=7.34 Hz, 2H), 7.48 (d, J=7.34 Hz,2H).

Step-3: Synthesis of (S)-diphenyl(pyrrolidin-2-yl)methanol (4)

To a stirred solution of 3 (0.3 g, 0.84 mmol) in dioxane (2 mL) at 0° C.was added 4M HCl in dioxane (2 mL) and the reaction mixture allowed towarm to room temperature and stirred for a further 2 h. The progress ofthe reaction was monitored by TLC for complete consumption of compound3. All volatiles were removed under reduced pressure to afford a residuethat was triturated with pentane to get 4 as an off-white solid (0.12 g,crude). LCMS: 254.01 (M+H). ¹H NMR (400 MHz, DMSO-d₆) δ 1.83-1.86 (m,4H), 3.08-3.17 (m, 2H), 4.86-4.88 (m, 1H), 6.58 (s, 1H), 7.20-7.27 (m,2H), 7.30-7.40 (m, 4H), 7.53 (d, J=7.82 Hz, 2H), 7.61 (d, J=7.82 Hz,2H), 8.09 (br s, 1H).

Step-4: Synthesis of tert-butyl(S)-(4-(2-(hydroxydiphenylmethyl)pyrrolidin-1-yl)-4-oxobutyl)carbamate(5)

A stirred solution of 4-((tert-butoxycarbonyl)amino)butanoic acid (0.12g, 0.62 mmol) and triethyl amine (0.2 mL, 2.07 mmol) in DCM (20 mL) at0° C. was treated with HOBt (0.08 g, 0.49 mmol) and EDC.HCl (0.1 g, 0.62mmol) and stirred for 5 min. To this reaction mixture was added compound4 (0.25 g, 1.22 mmol) and reaction mixture allowed to warm to roomtemperature and stirred for a further 16 h. The reaction was monitoredby TLC for complete consumption of compound 4. The reaction mixture wasdiluted with water (2 mL) and extracted with DCM (5 mL×2). The combinedorganic layer was washed with water (2 mL), dried over anhydrous sodiumsulfate and concentrated under reduced pressure to afford crude. Thecrude product was purified by silica gel column chromatography (230-400mesh) using 35% EtOAc in hexanes to afford 5 as a colourless solid (0.1g, 60% yield). LCMS: 439 (M+H).

Step-5: Synthesis of(S)-4-amino-1-(2-(hydroxydiphenylmethyl)pyrrolidin-1-yl)butan-1-one (6)

To a stirred solution of 5 (0.1 g, 0.23 mmol) in dioxane (2 mL) at 0° C.was added 4M HCl in dioxane (2 mL) and the reaction mixture allowed towarm to room temperature and stirred for an additional 2 h. The progressof the reaction was monitored by TLC for complete consumption ofcompound 5. All volatiles were removed under reduced pressure to afforda residue that was triturated with pentane to provide 6 as an off-whitesolid (0.09 g, crude). LCMS: 339.15 (M+H).

Step-6: Synthesis of ethyl(S)-(4-(2-(hydroxydiphenylmethyl)pyrrolidin-1-yl)-4-oxobutyl)glycinate(7)

To a stirred solution of compound 6 (0.09 g, 0.24 mmol) in DCE (20 mL)was added ethyl 2-oxoacetate (50% solution in toluene, 0.03 g, 0.28mmol) and stirred at room temperature for 30 min. To this was added STAB(0.2 g, 0.96 mmol) and TFA (0.03 g, 0.24 mmol) to the reaction mixtureat room temperature and reaction mixture was further stirred for 16 h.The reaction was monitored by TLC for complete consumption of compound6. The reaction mixture was quenched with saturated NaHCO₃ solution (2mL) and extracted with DCM (5 mL×2). The combined organic layer waswashed with water (2 mL) and brine (2 mL) respectively then dried overanhydrous sodium sulfate and concentrated under reduced pressure toafford compound 7. The crude product was purified by silica gel columnchromatography (100-200 mesh) using 15% EtOAc in hexanes to afford 7 asan off white solid (0.06 g, 60% yield). LCMS: 425.2 (M+H). ¹H NMR (400MHz, CHLOROFORM-d) δ 0.85-1.02 (m, 1H), 1.24-1.31 (t, J=7.14 Hz, 3H),1.53-1.58 (m, 4H), 1.72-1.80 (m, 1H), 1.93-2.01 (m, 1H), 2.03-2.07 (m,1H), 2.31-2.39 (m, 2H), 2.54-2.64 (m, 2H), 2.89-3.00 (m, 1H), 3.37 (s,2H), 4.18 (q, J=6.89 Hz, 2H), 5.14-5.17 (m, 1H), 5.28 (s, 1H), 7.34-7.42(m, 3H), (remaining protons were obscured by solvent peaks).

Step-7: Synthesis of(5)-1-(4-(2-(hydroxydiphenylmethyl)pyrrolidin-1-yl)-4-oxobutyl)imidazolidine-2,4-dione

To a stirred solution of compound 7 (0.06 g, 0.12 mmol) in AcOH (5 mL)was added KOCN (0.02 g, 0.25 mmol) and the reaction mixture was heatedat 100° C. for 2 h. The progress of the reaction was monitored by TLC.Reaction mixture was concentrated under reduced pressure and residue wastaken in saturated solution of NaHCO₃ solution (5 mL) and extracted withEtOAc (5 mL×2). The organic extract was washed with water (5 mL) andbrine (5 mL) respectively then dried over anhydrous sodium sulfate andconcentrated under reduced pressure to dryness. The residue was purifiedby combiflash chromatography (eluting with 80-90% EtOAc in hexane toafford the final product as a colourless solid (12 mg, 23%). ¹H NMR (400MHz, DMSO-d₆) δ 1.32-1.43 (m, 1H), 1.57-1.59 (m, 3H), 1.81-1.93 (m, 2H),1.99-2.09 (m, 2H), 3.54-3.67 (m, 1H), 3.82 (s, 2H), 4.98-5.13 (m, 1H),6.44-6.63 (m, 1H), 7.17-7.28 (m, 4H), 7.29-7.34 (m, 3H), 7.42-7.44 (m,2H), 10.44 (br s, 1H), (remaining protons were obscured by solventpeaks). LCMS: 420.15 (M−H). HPLC purity: 93.17%.

Example 80: Synthesis of3-((3-((2-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)pyrrolidin-1-yl)sulfonyl)propyl)amino)-6-methoxypyridin-2(1H)-one(alternate ID: 0081)

Step-1: Synthesis of tert-butyl2-(methoxy(methyl)carbamoyl)pyrrolidine-1-carboxylate (9)

To a mixture of 8 (10.0 g, 4.64 mmol) in dry DCM (100 mL),N,O-dimethylhydroxylamine (4.98 g, 5.11 mmol), NMM (5.16 g, 5.11 mmol)and EDC (9.76 g, 5.11 mmol) were added at −5° C. and the reactionmixture was stirred at room temperature for 3 h. The progress of thereaction was monitored by TLC. After completion of the reaction, thereaction mixture was quenched with 1N HCl solution and extracted withDCM. The combined organic layer were washed with brine, dried overanhydrous Na₂SO₄ and concentrated under reduced pressure to afford 9.Yield: 7.0 g, 58%; NMR: ¹H NMR (400 MHz, DMSO-d₆) δ 4.61-4.50 (m, 1H),3.69 (d, J=11.1 Hz, 3H), 3.40-3.30 (m, 2H), 3.10 (d, J=6.5 Hz, 3H),2.18-2.15 (m, 1H), 1.78-1.75 (m, 3H), 1.35 (d, J=25.4 Hz, 9H).

Step-2: Synthesis of tert-butyl2-(3-(cyclopropylmethoxy)-4-fluorobenzoyl)pyrrolidine-1-carboxylate (11)

To a stirred solution of 10 (4.0 g, 16.0 mmol) in dry THF (40 mL),activated Mg (0.768 g, 32.0 mmol) and 12 (1 pinch) was added followed byaddition of CH₃I (0.3 mL) was added and stirred at 60° C. for 30 min. 9(4.03 g, 16.0 mmol) in THF (40 mL) was added and reaction was stirred at60° C. for 12 h. The progress of the reaction was monitored by TLC.After completion of the reaction, the reaction mixture was quenched withwater and extracted with EtOAC. The combined organic layer were washedwith brine, dried over anhydrous Na₂SO₄ and concentrated under reducedpressure. The crude product was purified by column chromatography using6% EtOAc/hexane to afford 11. Yield: 3 g, 52%; NMR: ¹H NMR (400 MHz,Chloroform-d3) δ 7.65-7.47 (m, 2H), 7.19-7.09 (m, 1H), 5.31-5.10 (m,1H), 3.93 (t, J=8.0 Hz, 1H), 3.70-3.41 (m, 1H), 2.31-2.29 (m, 1H),1.92-1.90 (m, 2H), 1.25 (d, J=4.0 Hz, 9H), 0.89-0.86 (m, 1H), 0.67 (q,J=6.8, 6.0 Hz, 2H), 0.37 (t, J=5.8 Hz, 2H) (remaining protons obscuredby solvent peaks).

Step-3: Synthesis of tert-butyl2-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)vinyl)pyrrolidine-1-carboxylate(12)

To a stirred solution of Ph₃PCH₃Br (5.29 g, 14.83 mmol) in dry THF (20mL), NaHMDS (1M in THF 14.5 mL, 14.83 mmol) was added at 0° C. and themixture then stirred at room temperature for 2 h. A solution of 11 (3.0g, 8.24 mmol) in THF (20 mL) was added dropwise at 0° C. and reactionwas stirred at room temperature for 12 h. The progress of the reactionwas monitored by TLC. After completion of the reaction, the reactionmixture was quenched with NH₄Cl solution and extracted with EtOAc. Thecombined organic layer were washed with brine, dried over anhydrousNa₂SO₄ and concentrated under reduced pressure. The crude product waspurified by column chromatography using 5% EtOAc/hexane to afford 12.Yield: 1.7 g, 49%; LCMS: 365.25 (M+1).

Step-4: Synthesis of tert-butyl2-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)pyrrolidine-1-carboxylate(13)

To a stirred solution of 12 (1.7 g, 4.66 mmol) in MeOH (20 mL), 10% Pd/C(0.7 g) was added and stirred under hydrogen atmosphere (balloonpressure) at room temperature for 2 h. The progress of the reaction wasmonitored by TLC. After completion of the reaction, the reaction mixturewas filtered through celite and filtrate was evaporated under reducedpressure to afford 13. Yield: 1.55 g, 92%; NMR: ¹H NMR (400 MHz,DMSO-d₆) δ 7.13 (td, J=11.7, 8.1 Hz, 1H), 6.84-6.68 (m, 2H), 3.86 (d,J=6.5 Hz, 2H), 3.09-3.02 (m, 1H), 1.81-1.63 (m, 2H), 1.53 (s, 1H),1.51-1.42 (m, 9H), 1.36 (dd, J=9.4, 6.6 Hz, 3H), 1.19-1.16 (m, 1H), 0.83(dd, J=9.7, 6.6 Hz, 1H), 0.58 (d, J=7.8 Hz, 2H), 0.31 (d, J=5.4 Hz, 2H).

Step-5: Synthesis of2-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)pyrrolidine (14)

To a stirred solution of 13 (1.55 g, 1.0 mmol) in 1:1 dioxane:MeOH (30mL) was added a 1M solution of, HCl in dioxane (15 mL) and the reactionmixture stirred at room temperature for 12 h. After completion of thereaction, the reaction mixture was concentrated and the residue waspurified by trituration with ether to afford 14. Yield: 1.3 g, crude;LCMS: 364.05 (M+1).

Step-6: Synthesis of ethyl3-((2-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)pyrrolidin-1-yl)sulfonyl)propanoate(16)

To a stirred solution of(R/S)-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)pyrrolidine 14(0.7 g, 2.60 mmol) in dry DCM (10 mL), Et₃N (1.1 mL, 7.9 mmol) was addedand stirred at room temperature for 10 min. A solution of 15 (0.64 g,3.18 mmol) in DCM (15 mL) was added dropwise at 0° C. and then stirredat room temperature for 16 h. The progress of the reaction was monitoredby TLC. After completion of the reaction, the reaction mixture wasquenched with water and extracted with DCM. The combined organic layerwere washed with brine, dried over anhydrous Na₂SO₄ and concentratedunder reduced pressure. The crude product was purified by columnchromatography using 20% EtOAC/hexane to afford 16. Yield: 0.4 g, 35.4%;LCMS: 428.05 (M+1).

Step-7: Synthesis of3-((2-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)pyrrolidin-1-yl)sulfonyl)propan-1-ol(17)

To a stirred solution of 16 (0.6 g, 1.40 mmol) in THF (15 mL), LiBH₄ (2Msolution in THF, 1.4 mL, 2.80 mmol) was added at 0° C. and then heatedto 60° C. and stirred for 4 h. The progress of the reaction wasmonitored by TLC. Upon completion the reaction was concentrated todryness under reduced pressure. The reaction mixture was diluted withwater and extracted with EtOAc. The combined organic layer were washedwith brine, dried over anhydrous Na₂SO₄ and concentrated under reducedpressure. The crude product was purified by column chromatography using30% EtOAc/hexane to afford 17. Yield: 0.375 g, 69%; LCMS; 386.05 (M+1).

Step-8 Synthesis of3-((2-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)pyrrolidin-1-yl)sulfonyl)propanal(18)

To a stirred solution of 17 (0.17 g, 0.44 mmol) in DCM (10 mL), PCC(0.257 g, 1.10 mmol) and molecular sieves (20 mg) were added at 0° C.and then stirred at room temperature for 3 h. The progress of thereaction was monitored by TLC. After completion of the reaction, thereaction mixture was filtered through celite and filtrate was evaporatedunder reduced pressure to afford 18. Yield: 0.075 g, crude: LCMS; 384.0(M+1).

Step-9: Synthesis of3-((3-((2-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)pyrrolidin-1-yl)sulfonyl)propyl)amino)-6-methoxypyridin-2(1H)-one

To a stirred solution of 18 (0.075 g, 0.19 mmol) in MeOH (50 mL), 7N NH₃in MeOH (6 drops) solution was added till pH=7 and reaction was allowedto stirred at room temperature. AcOH (4 drops) was added drop wise tillpH=6 followed by addition of 3-amino-6-methoxypyridin-2(1H)-one 19(0.027 g, 0.19 mmol) and stirred at room temperature for 10 min. Afterthat NaCNBH₃ (0.037 g, 0.58 mmol) was added at 0° C. The progress of thereaction was monitored by TLC. After completion of the reaction, thereaction mixture was quenched with NaHCO₃ solution and extracted withDCM. The combined organic layer were washed with brine, dried overanhydrous Na₂SO₄ and concentrated under reduced pressure. The crudeproduct was purified by column chromatography using 1-2% MeOH/DCM toafford the final product as a mixture of diastereoisomers. Yield: 0.01g, 10%; HPLC purity: 95.45%; LCMS: observed mass; 508.13 (M+1). ¹H NMR(400 MHz, DMSO-d₆) δ 7.16-7.05 (m, 1H), 7.03-6.91 (m, 1H), 6.84-6.76 (m,1H), 6.33 (s, 1H), 3.87 (td, J=9.9, 8.4, 4.5 Hz, 3H), 3.67 (d, J=1.5 Hz,2H), 3.31-3.22 (m, 1H), 3.18-2.89 (m, 8H), 1.96-1.88 (m, 2H), 1.69 (s,1H), 1.51 (s, 1H), 1.26-1.14 (m, 8H), 0.62-0.52 (m, 2H), 0.37-0.28 (m,2H).

Example 81 Synthesis of 3-amino-6-methoxypyridin-2(1H)-one (19) requiredfor the synthesis of3-((3-((2-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)pyrrolidin-1-yl)sulfonyl)propyl)amino)-6-methoxypyridin-2(1H)-one

Step-1: Synthesis of 6-methoxy-3-nitropyridin-2(1H)-one (21)

To a stirred solution of NaOtBu (12.43 g, 129.3 mmol) in liq. NH₃ (50mL) was added 20 (4 g, 25.9 mmol) followed by a solution of tert-butylhydroperoxide (3.06 mL) in THF (50 mL) at −78° C. The reaction mixturewas allowed to stir at room temperature for 12 h. Then the reactionmixture was diluted with water, neutralized with 1N HCl solution andextracted with DCM (100 mL×2). Combined organic layer was washed withbrine, dried over anhydrous Na₂SO₄ and concentrated under reducedpressure. The crude compound thus obtained was purified by columnchromatography using 30% EtOAc in hexane as eluent to afford 21. Yield:2.8 g, 63%; NMR: ¹H NMR (400 MHz, DMSO-d₆) δ 12.85 (s, 1H), 8.42 (d,J=8.9 Hz, 1H), 6.32-6.22 (m, 1H), 3.95 (s, 3H).

Step-2: Synthesis of 3-amino-6-methoxypyridin-2(1H)-one (19)

To a stirred solution of 21 (0.7 g, 4.11 mmol) in MeOH (7 mL) were added10% Pd/C (0.25 g) and glacial acetic acid (catalytic) and the reactionmixture was stirred at room temperature for 2 h under hydrogen balloonpressure. Then the reaction mixture was filtered through celite andfiltrate was evaporated under reduced pressure to afford 19 (0.54 g,crude) which was used as such in next step without further purification.

Example 82 Synthesis of3-((3-((2-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)pyrrolidin-1-yl)sulfonyl)propyl)amino)piperidine-2,6-dione(alternate ID: 0012)

To a stirred solution of 22 (0.042 g, 0.26 mmol) in MeOH (50 mL), 7N NH₃(10 drops) in MeOH solution was added till pH=7 and reaction was allowedto stirred at room temperature. AcOH (5 drops) was added till pH=6followed by addition of 18 (0.1 g, 0.26 mmol) and stirred at roomtemperature for 10 min. After that NaCNBH₃ (0.049 g, 0.78 mmol) wasadded at 0° C. and stirred at room temperature for 12 h. The progress ofthe reaction was monitored by TLC. After completion of the reaction, thereaction mixture was quenched with NaHCO₃ solution and extracted withDCM. The combined organic layer were washed with brine, dried overanhydrous Na₂SO₄ and concentrated under reduced pressure. The crudeproduct was purified by column chromatography using 1-5% MeOH/DCM toafford3-((3-((2-(1-(3-(cyclopropylmethoxy)-4-fluorophenyl)ethyl)pyrrolidin-1-yl)sulfonyl)propyl)amino)piperidine-2,6-dione.Yield: 0.045 g, 35%; HPLC purity: 94.37%; LCMS: observed mass; 496.15(M+1). ¹H NMR (400 MHz, DMSO-d₆) δ 10.69 (s, 1H), 7.13-7.11 (m, 1H),7.04-6.92 (m, 1H), 6.82-6.80 (m, 1H), 4.00-3.87 (m, 3H), 3.27 (dd,J=12.5, 5.4 Hz, 1H), 3.13 (q, J=7.5 Hz, 2H), 3.02-2.87 (m, 2H),2.82-2.61 (m, 3H), 2.54 (d, J=6.1 Hz, 1H), 2.03-2.01 (m, 1H), 1.75-1.72(m, 6H), 1.57-1.47 (m, 1H), 1.35-1.16 (m, 4H), 0.83 (dd, J=10.2, 6.8 Hz,2H), 0.62-0.52 (m, 2H), 0.37-0.28 (m, 2H).

Biological Methods

A. Drugs, Reagents and Cell Lines

Test compounds are suspended in DMSO at a concentration, e.g., of 100mmol/L, fluorodeoxyuridine (FUdR) that can be obtained from Sigma (StLouis, Mo.) and maintained in sterile double-distilled water at stockconcentrations of 50 mmol/L.

Recombinant human deoxyuridine nucleotidohydrolase (dUTPase) isexpressed and purified as described in Ladner R D, Carr S A, HuddlestonM J, McNulty D E, Caradonna S J. J Biol Chem. 1996 Mar. 29;271(13):7752-7. All drugs stocks are aliquoted and diluted asappropriate prior to use. The oligonucleotide primer, templates andfluorophore- and quencher-labeled detection probes are synthesized byIntegrated DNA Technologies (Coralville, Iowa), subjected topolyacrylamide gel electrophoresis purification and reconstituted inOmnipur sterile nuclease-free water (EMD Chemicals USA, Gibbstown N.J.)at a stock concentration of 100 μmol/L. The two non-emissive (dark)quenching molecules incorporated into the detection probes include theIowa black fluorescein quencher (IBFQ; absorption max 531 nm) and ZEN(non-abbreviation; absorption max 532 nm). The fluorescent labelutilized is 6-FAM (5′-carboxyfluorescein; excitation max.=494 nm,emission max.=520 nm). Probes are further diluted to a working stock of10 μmol/L and aliquoted to avoid repeated freeze/thaw cycles. AmpliTaqGold DNA Polymerase, GeneAmp 10×PCR Buffer 2, MgCl₂ and MicroAmp Optical96-well Reaction Plates are purchased from Applied Biosystems (Carlsbad,Calif.). dNTPs are purchased individually at stock concentrations of 100mmol/L from New England Biolabs at HPLC-certified >99% purity (Ipswich,Mass.).

B. Assay Components, Instrumentation and Real-Time FluorescenceConditions

Reaction mixtures contained primer, probe and template at an equimolarfinal concentration of 0.4 μmol/L. Magnesium chloride (MgCl₂) isincluded at a final concentration of 2 mmol/L. Non-limiting dNTPs areincluded in the reaction mix in excess at a final concentration of 100μmol/L (dUTP/dTTP is excluded). AmpliTaq Gold DNA polymerase is added at0.875 U/reaction, 2.5 μl of 10×PCR buffer 2 added and nuclease-freeddH₂O added to a final reaction volume of 25 μl. For dUTP inhibitionanalysis, the volume of ddH₂O is further modified to accommodate anadditional 1 μl of dUTPase (10 ng/μl) and 1 μl of inhibitor or DMSOcontrol. Thermal profiling and fluorescence detection is performed usinga customized thermal program on board a Roche Lightcycler LC480Instrument II. For analysis of dNTPs, the thermal profile consisted ofan 8 min 37° C. step followed by a 10 min 95° C. step to ‘hot-start’ theTaq polymerase and a primer extension time of up to 30 min at 60° C. Rawfluorescence spectra for 6-FAM is measured at specified time intervalsto follow assay progression on board a Roche LightCycler 480 SoftwareVersion 1.5 and exported and analyzed in Microsoft Excel (Microsoft,Redmond Wash.) and Prism (GraphPad Software, La Jolla Calif.).Fluorescence values for blank reactions (limiting dNTP omitted) aresubtracted to give normalized fluorescence units (NFU) to account forbackground fluorescence.

C. MTS Growth Inhibition Assay

The Cell Titer AQueous MTS assay (Promega) is carried out according tothe manufacturers guidelines. IC_(50(72 h)) values are calculated fromsigmoidal-dose response curves utilizing Prism (Graphpad, San Diego,Calif.). The combination effect is determined by the combination index(CI) method utilizing Calcusyn software (Biosoft, Ferguson, Mo.).Fraction affected (FA) is calculated from the percent growth inhibition:FA=(100−% growth inhibition)/100. CI values <1, synergism; 1-1.2,additive and >1.2, antagonism.

D. Colony Formation Assay

Colony forming assay showing the ability of colon (SW620, HCT116),non-small cell lung (A549, H460, H1299 and H358) and breast (MCF7)cancer cells to survive and proliferate following transient 24 hourexposure to test compounds, FUdR and combinations are determined.Specifically, cells are seeded at densities between 50 and 100cells/well in 24-well plates. Twenty-four hours later, cells are treatedwith increasing concentrations of a rtest compound, a fixed dose of FUdRand combinations of these. After 24 hours, drug is removed, cells arerinsed and allowed to outgrow for 10-14 days. At the conclusion of theoutgrowth, cells are fixed in 60% ice cold methanol and stained with0.1% crystal violet, scanned and counted. Data is presented aspercentage of untreated controls (mean±SD). Fraction affected andcombination indexes are calculated according to the method of Chou andTalalay where <1 is indicative of a synergistic drug interaction.

E. In Vivo Analysis

Xenograft experiments are conducted in male NU/NU nude mice (CharlesRiver, Wilmington, Mass.) that are 6-8 weeks old. Subcutaneous A549xenografts are established and allowed to grow until they reached ˜50mm³ (day 1). Animals are randomized to treatment groups: vehicle,pemetrexed 50 mg/kg, a test compound and combination of pemetrexed plusa test compound (n=5, group). Pemetrexed is administered at 50 mg/kg byintraperitoneal injection every two days. Test compound is administered,e.g., at 75 mg/kg by intraperitoneal injection every two days. Thecombination of pemetrexed and the test compound is administered byintraperitoneal injection, e.g., every two days. Two perpendiculardiameters of tumors are measured every 2 days with a digital caliper bythe same investigator. Tumor volume is calculated according to thefollowing formula: TV (mm³)=(length[mm]×(width[mm]²)/2. Mice areinspected every day for overall health and bodyweight is measured every2 days as an index of toxicity. All animal protocols are approved by theUSC Institutional Animal Care and Use Committee (IACUC).

F. dUTPase Inhibition

Test compounds are screened in a fluorescence-based assay. The assayemploys a DNA polymerase-based approach utilizing an oligonucleotidetemplate with 3 distinct regions: a 3′ primer binding region, amid-template dUTP/thymidine triphosphate (TTP) detection region and a 5′6-Flavin adenine mononucleotide (FAM)-labeled probe binding region thatincorporates a black hole quenching moiety. During the reaction, theprobe and primer hybridize to the oligonucleotide template to form thetemplate:primer:probe complex. When Taq polymerase binds to the primerin the TPP complex and dUTP is present, successful extension of thenascent strand occurs and the inherent 5′ to 3′ exonuclease activity ofTaq polymerase cleaves and displaces the 6-FAM-labeled probe in a 5′ to3′ direction, releasing the 6-FAM fluorophore from its proximity to thethree quenchers. This displacement effectively disrupts the Försterresonance energy transfer (FRET) and the resulting fluorescence detectedupon excitation is directly proportional to the amount of the dUTPavailable in the assay for incorporation. Conversely, when the dUTP isunavailable, exhausted, or degraded by dUTPase and is no longeravailable for incorporation, Taq polymerase stalls and extension delayand/or chain termination of the nascent strand occurs. In this instance,probe hydrolysis/degradation does not occur and the probe remains darkas fluorescence remains quenched via FRET. Since fluorescence isdirectly proportional to the concentration of dUTP, the assay is easilymodified to measure dUTP and the effects of inhibitors on dUTPhydrolysis by the enzyme dUTPase. The template BHQ-DT6 (Black HoleQuencher-Detection Template 6) for detecting up to 60 pmols of dUTP isincluded for this application of the assay along with 50 pmols of dUTPand 5 ng of recombinant dUTPase. The reaction is incubated at 37° C. for8 mins and terminated by a 10 min incubation at 95° C. to simultaneouslyinactivate dUTPase and activate the hot-start Taq polymerase. Thefluorescence generated during the detection step is directlyproportional to the concentration of dUTP remaining after the 8 minincubation. The concentration of dUTP at reaction termination andtherefore inhibition of dUTPase in the presence and absence ofinhibitors and appropriate dimethyl sulfoxide (DMSO) controls can bedetermined.

Example alternate dUTPase % relative inhibition No. ID isomer IC₅₀ (μM)vs. positive control 79 1229 1 75 80 81 mixture of 22 diastereomers 8212 mixture of 13 diastereomers

Percentage of relative inhibition is a straightforward comparison of theinhibitory activity of a candidate compound relative to that of thepositive control compound across three molar concentrations. Todetermine % relative inhibition, candidate compounds were screened fordUTPase enzyme inhibitory activity at three descending concentrations,typically, 100, 50 and 25 μmon using the methodology described. Thepercentage of dUTPase enzyme inhibition for each candidate compound ateach of the concentrations was calculated and directly compared to thatof the in-run positive control at the equivalent concentrations. Theresulting percentage inhibition at each concentration was subsequentlyaveraged to give a mean percentage inhibition indicative of thecandidate compounds performance across the three molar concentrationstested when compared to the positive control.

Test compounds are evaluated for their antitumor activity in colorectalcancer cells using the MTS growth inhibition assay. HCT116 and SW620cells are exposed to increasing concentrations of each agent for 72hours and growth inhibition is directly compared to vehicle-treatedcontrols. The NSCLC cell lines A549 and H1299 are exposed to increasingconcentrations of each agent for 72 hours and growth inhibition isdirectly compared to vehicle-treated controls.

G. Growth Inhibition

MTS growth inhibition assays are performed to evaluate the effectivenessof the test compounds alone and in combination with the fluoropyrimidinethymidylate synthase (TS) inhibitor 5-fluorouracil (5-FU) at inhibitingthe growth of colorectal (HCT116 and SW620) cell line models. Increasingconcentrations of 5-FU between 0 and 100 μmol/L demonstrateddose-dependent increases in growth inhibition in both the colorectalcancer cell lines evaluated. Simultaneous treatment with increasingconcentrations of 5-FU and a test compound at fixed concentrations of 25μmol/L is determined.

H. Reducing Cancer Cell Viability

Colony forming assays are performed to evaluate the effectiveness oftest compounds alone and in combination with the fluoropyrimidinethymidylate synthase (TS) inhibitor fluorodeoxyuridine (FUdR) atreducing cancer cell viability in colorectal (HCT116), breast (MCF-7)and non-small cell lung (H1299, A549, H358 and H460) cell line models.Increasing concentrations of FUdR between 0.5 and 2.5 μmol/Ldemonstrated dose-dependent decreases in colonies formed in all celllines evaluated. In colorectal cancer cells, concentrations of testcompounds ranging e.g., from 3.1 μmol/L to 50 μmol/L are combined with0.5 μmol/L FUdR in HCT116 cells and 1 μmol/L FUdR in SW620 cells.

It should be understood that although the present invention has beenspecifically disclosed by certain aspects, embodiments, and optionalfeatures, modification, improvement and variation of such aspects,embodiments, and optional features can be resorted to by those skilledin the art, and that such modifications, improvements and variations areconsidered to be within the scope of this disclosure.

The invention has been described broadly and generically herein. Each ofthe narrower species and subgeneric groupings falling within the genericdisclosure also form part of the invention. In addition, where featuresor aspects of the invention are described in terms of Markush groups,those skilled in the art will recognize that the invention is alsothereby described in terms of any individual member or subgroup ofmembers of the Markush group.

What is claimed is:
 1. A compound of Formula (IA):

or a tautomer thereof, or a pharmaceutically acceptable salt of each ofthe foregoing, or a pharmaceutically acceptable solvate of each of theabove mentioned, wherein A is

L¹ is

wherein one or more hydrogens are optionally substituted with C₁-C₃alkyl and/or two geminal hydrogens together with the carbon to whichthey are attached are optionally replaced with a 3-5 memberedheterocyclyl or a 3-5 membered cycloalkyl; and wherein p is 0, 1, 2, 3,4, or 5 and z is 0, 1, 2, 3, 4, or 5; L² is —C(O)— or —S(O)₂—; thenitrogen containing ring attached to L² and L³ is optionallysubstituted;

is selected from the group consisting of:

L³ is a bond or —C(R³⁰⁰)₂—; each R³⁰⁰ independently is a hydrogen,hydroxyl, an optionally substituted C₁-C₆ alkyl, or an optionallysubstituted phenyl; and B is an optionally substituted 6-10 memberedaryl; an optionally substituted 5-15 membered heteroaryl; an optionallysubstituted 4-15 membered heterocyclyl; or an optionally substituted3-15 membered cycloalkyl, or a B substituent together with L² or L³,form a 5-7 membered cycloalkyl or a heterocyclyl.
 2. The compound ofclaim 1, wherein the compound is of Formula (I):

wherein the variables are defined as in formula (IA) above in claim 1.3. The compound of claim 1, wherein B is selected from the groupconsisting of:

wherein each R⁶ independently is hydrogen, an optionally substitutedC₁-C₆ alkoxy, or halo; each R⁷ independently is an optionallysubstituted C₁-C₆ alkyl, an optionally substituted C₂-C₆ alkenyl, anoptionally substituted C₂-C₆ alkynyl, an optionally substituted C₃-C₈cycloalkyl, an optionally substituted C₃-C₁₀ heteroaryl, an optionallysubstituted C₃-C₁₀ heterocyclyl, or an optionally substituted C₆-C₁₀aryl; or R⁶ and R⁷ together with the atoms they are attached to form anoptionally substituted 5-7 membered ring; or 2 R⁶ groups together withthe atoms they are attached to form an optionally substituted 5-7membered ring; each R⁶¹ and R⁶² is independently N or CH, provided thatat least one of R⁶¹ and R⁶² is N, each R⁶³ is independently NR⁹⁰, S, orO; each R⁶⁴ is independently N or CH; each R⁹⁰ is independently hydrogenor R⁷; each R¹-R³ independently is H, halo, an optionally substitutedC₁-C₆ alkyl, an optionally substituted 4-15 membered heterocyclyl, or—OR²⁰ or, if two of R¹-R³ are on adjacent carbon atoms, then two suchsubstituents together with the atoms they are attached to form anoptionally substituted 5-7 membered ring; R²⁰ is (CH₂)_(w)—R²¹, anoptionally substituted C₃-C₆ cycloalkyl, or an optionally substitutedC₁-C₆ alkyl; R²¹ is an optionally substituted C₃-C₆ cycloalkyl, anoptionally substituted C₆-C₁₀ aryl, an optionally substituted 5-15membered heteroaryl, an optionally substituted 4-15 memberedheterocyclyl, an optionally substituted C₁-C₁₀ alkyl, an optionallysubstituted C₂-C₁₀ alkenyl, an optionally substituted C₂-C₁₀ alkynyl, anoptionally substituted 4-15 membered heterocyclyl, or

wherein each R²²-R²⁴ independently is an optionally substituted C₁-C₃alkyl or hydroxy or two of R²²-R²⁴ together with the carbon atoms theyare attached to form a 3-7 membered ring; and w is 1, 2, 3, 4, or 5; orB is

wherein each R¹-R³ independently is H, halo, an optionally substitutedC₁-C₆ alkyl, an optionally substituted 4-15 membered heterocyclyl; or R¹and R² together with the atoms they are attached to form an optionallysubstituted 5-7 membered ring; or R² and R³ together with the atoms theyare attached to form an optionally substituted 5-7 membered ring.
 4. Thecompound of claim 1, wherein L¹ is selected from the group consistingof:

wherein the left side of the moieties are attached to A.
 5. The compoundof claim 1, wherein L³ is a bond.
 6. The compound of claim 1, wherein L³is —C(R³⁰⁰)₂—.
 7. The compound of claim 1, wherein B is

R²⁰ is (CH₂)_(w)—R²¹, an optionally substituted C₃-C₆ cycloalkyl, or anoptionally substituted C₁-C₆ alkyl; R²¹ is an optionally substitutedC₃-C₆ cycloalkyl, an optionally substituted C₆-C₁₀ aryl, an optionallysubstituted 5-15 membered heteroaryl, an optionally substituted 4-15membered heterocyclyl, an optionally substituted C₁-C₁₀ alkyl, anoptionally substituted C₂-C₁₀ alkenyl, an optionally substituted C₂-C₁₀alkynyl, an optionally substituted 4-15 membered heterocyclyl, or

wherein each R²²-R²⁴ independently is an optionally substituted C₁-C₃alkyl or hydroxy or two of R²²-R²⁴ together with the carbon atoms theyare attached to form a 3-7 membered ring; and w is 1, 2, 3, 4, or 5; orB is selected from the group consisting of:

wherein the alkoxy group is further optionally substituted wherein 1-5hydrogen atoms are optionally substituted; and R⁷⁰ is hydrogen or anoptionally substituted C₁-C₁₀ alkyl.
 8. A composition comprising acompound of claim 1 and at least one pharmaceutically acceptableexcipient or carrier.
 9. A method of inhibiting dUTPase, wherein themethod comprises contacting the dUTPase with an effective amount of thecompound of claim
 1. 10. A method of inhibiting the growth of a cancercell comprising contacting the cell with a therapeutically effectiveamount of the compound of claim 1 and a therapeutically effective amountof a dUTPase directed therapeutic, thereby inhibiting the growth of thecancer cell; wherein the cancer cell is selected from a colorectalcancer cell, a breast cancer cell, or a non-small cell lung cancer cell.11. The method of claim 10, wherein the dUTPase-directed therapeutic isselected from fluoropyrimidines and antifolates.
 12. A method oftreating cancer in a patient whose treatment is impeded by theexpression or overexpression of dUTPase, comprising administering atherapeutically effective amount of the compound of claim 1 to thepatient, wherein the cancer is colorectal cancer, breast cancer, ornon-small cell lung cancer.
 13. The method of claim 12, furthercomprising detecting overexpression of dUTPase in a cell or tissuesample isolated from the patient.
 14. A method of treating colorectal,breast, or non-small cell lung cancer in a patient selected fortreatment by overexpression of dUTPase in a sample isolated from thepatient, comprising administering to the patient a therapeuticallyeffective amount of the compound of claim
 1. 15. A kit comprising acompound of claim 1, and instructions for diagnostic or therapeutic useas described herein.